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Rapid and precise detection of harmful substances in food products is essential for ensuring public health and safety. This study introduces a novel surface-enhanced Raman spectroscopy (SERS) substrate, composed of a molybdenum disulfide­silver nanocomposite, applied to flexible, water-resistant filter paper for detecting melamine and bisphenol A (BPA) in milk. Optimized molybdenum disulfide (NMS) nanoflowers (NFs) were synthesized through hydrothermal methods and high-temperature annealing, then modified with silver (Ag) nanoparticles to form the NMS-Ag nanocomposite (NMSA6). This substrate greatly enhances the Raman signal, achieving an enhancement factor of approximately 1.49 × 107 and a detection limit as low as 10-11 M for simultaneous multi-component analysis. Finite-difference time-domain (FDTD) simulations confirm the enhancement mechanism. The NMSA6 substrate demonstrates remarkably low detection limits for BPA and melamine, facilitating the analysis of various hazardous substances. These findings highlight the substrate's potential for highly sensitive, label-free detection, presenting a viable tool for food safety monitoring.

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The paper highlights how significant characteristics of liver can be modeled in tissue-engineered constructs using unconventional scaffolds. Hepatic lobular organization and metabolic zonation can be mimicked with decellularized plant structures with vasculature resembling a native-hepatic lobule vascular arrangement or silk blend scaffolds meticulously designed for guided cellular arrangement as hepatic patches or metabolic activities. The functionality of hepatocytes can be enhanced and maintained for long periods in naturally fibrous structures paving way for bioartificial liver development. The phase I enzymatic activity in hepatic models can be raised exploiting the microfibrillar structure of paper to allow cellular stacking creating hypoxic conditions to induce in vivo-like xenobiotic metabolism. Lastly, the paper introduces amalgamation of carbon-based nanomaterials into existing scaffolds in liver tissue engineering.

Unconventional scaffolds have the potential to meet the current challenges in liver tissue engineering- loss of hepatic morphology and functions over long-term culture, absence of native-like cell-cell and cell-matrix interactions, organization of hepatocytes into lobular structures exhibiting metabolic variations-which hinder pharmaceutical analysis, regenerative therapies and artificial organ development. Paper with cellulose microfibril network develops cellular aggregates with hypoxic conditions that influence enzymes of xenobiotic metabolism proving to be a better scaffold for hepatotoxicity testing compared with conventional monolayers in tissue culture plates. Decellularized plant stems provide already-built vasculature to be exploited for the development of intricate vessel networks that exist in hepatic lobules aiding in regenerative medicine for hepatic pathologies. Fibrous plant structures are excellent materials for the immobilization of hepatocytes and improve albumin secretion enabling their use in bioartificial liver development. Biomimicry of metabolic zonation in hepatic lobules can be achieved with perfusion culture using silk blend scaffolds with varying proportions of the liver matrix that orchestrate cellular function. The mechanical properties of silk allow the fabrication of structures that resemble liver anatomy to generate native-like hepatic lobules. Nanomaterials have immense potential as a component of composite material development for scaffolds to achieve improved predictive ability in pharmacokinetics. Most of these unconventional scaffolds have the added advantage of being readily available, accessible, affordable and sustainable for liver tissue engineering applications. Conclusively, the shift of attention away from conventional scaffolds poses a promising future in the field of tissue engineering.

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Residual chlorine from widespread disinfection processes forms byproducts in water that are harmful to humans and ecosystems. Portable sensors are essential tools for the on-site monitoring of residual chlorine in environmental samples. Here, an inexpensive colorimetric sensor was developed by grafting via amidation the chromogen orthotolidine (OTO) to the surface of a TEMPO-oxidized cellulose filter paper (O-TOFP). A thorough characterization of the sensor strip demonstrated that it was highly stable and that it could be stored for a long period before usage. O-TOFP had a fast response time of 30 s, was highly selective for residual chlorine ions (ClO-) with an accuracy of at least 95 %, and exhibited an excellent limit of detection of only 0.045 mg/L when combined with smartphone image acquisition. With its many positive features, the easy-to-use and robust O-TOFP sensor described here could become a useful tool for the determination of residual chlorine in different water samples.

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An amino functionalized paper-based material that utilized amino functionalized polymer particles as sensing probes and adsorption sites was fabricated via internal sizing technology for application in formaldehyde detection and adsorption. A large specific surface area and the porous structure of the paper fibers enable the application of the composite paper-based material as a sensor at low concentrations of primary amine groups. The material reacts with low levels of formaldehyde, resulting in a concentration-based change in the pH, which is rapidly expressed as a color change. After exposure to formaldehyde (0.02 mg/m3) for 10 min, the color of the composite paper-based material changed from pink to brown, demonstrating the high sensitivity of the material, and this transition could be clearly observed using the naked eye. Additionally, the composite paper-based material acts as an adsorbent at a high content of amino groups, owing to a rapid addition reaction with formaldehyde, exhibiting a high adsorption capacity. Considering the high sensitivity, adsorption capacity, and adsorption speed for formaldehyde, the as-developed composite paper-based material exhibits promising application potential in the field of formaldehyde detection and adsorption.

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Tibetan strawberry (Fragaria nubicola) is a wild medicinal and edible plant in Tibet possessing various health benefits such as neuroprotection and anti-oxidation. However, there has been little study reported on its chemical constituents. To investigate the inhibitors of monoamine oxidase B (MAO-B) in Tibetan strawberry, we immobilized the enzyme onto cellulose filter paper for the first time to develop a new screening method. Two known glycosides (compounds 1 and 2) and one new iridoid glucoside (Compound 3) were fished out by this method, which was found to effectively inhibit MAO-B with IC50 values of 16.95 ± 0.93, 24.69 ± 0.20, and 46.77 ± 0.78 µM, respectively. Molecular docking and kinetic analysis were performed to reveal the inhibition mechanism of these compounds. Furthermore, compound 1 exhibited neuroprotective effects against 6-OHDA-induced injury on PC12 cells. The developed method exhibits the advantages of rapidness and effectiveness in screening of MAO-B inhibitors from complex herbal extracts.

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Harvesting energy from air water (atmospheric moisture) promises a sustainable self-powered system without any restrictions from specific environmental requirements (e.g., solar cells, hydroelectric, or thermoelectric devices). However, the present moisture-induced power devices traditionally generate intermittent or bursts of energy, especially for much lower current outputs (generally keeping at nA or µA levels) from the ambient environment, typically suffering from inferior ionic conductivity and poor hierarchical structure design for manipulating sustained air water and ion-charge transport. Here, we demonstrate a universal strategy to design a high-performance bilayer polyelectrolyte ion paper conductor for generating continuous electric power from ambient humidity. The generator can produce a continuous voltage of up to 0.74 V and also an exceptional current of 5.63 mA across a single 1.0 mm-thick ion paper conductor. We discover that the sandwiched LiCl-nanocellulose-engineered paper promises an ion-transport junction between the negatively and positively charged bilayer polyelectrolytes for application in MEGs with both high voltage and high current outputs. Moreover, we demonstrated the universality of this bilayer sandwich nanocellulose-salt engineering strategy with other anions and cations, exhibiting similar power generation ability, indicating that it could be the next generation of sustainable MEGs with low cost, easier operation, and high performance.

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The main mechanism that causes resistance to carbapenem, one of the most potent antibiotic available, in Enterobacterales bacterial isolates, is due to Klebsiella pneumoniae carbapenemase (KPC) production by the bacterium. KPC is spread worldwide, requiring laboratories to be capable of identifying this enzyme, however some methods can be expensive for small laboratories, especially in developing countries. Therefore, the development of methods with low cost of reagents for the detection of KPC enzyme is necessary. The objective of this study was to evaluate the detection of KPC enzyme by MALDI-TOF MS from inactivated bacteria impregnated in filter paper. A total of 129 Enterobacterales isolates were impregnated in filter paper, and after 7 days at room temperature, they were subjected to a protein extraction protocol and spectra acquisition, in triplicates, by MALDI-TOF MS. The spectra were evaluated and KPC was identified according to the presence of a peak of 28,712.62 ± 27.80 m/z. Considering the presence of the KPC peak in at least one spectrum of the triplicates, this method presented 60.8% sensitivity and 96.4% specificity. However, considering the presence of KPC peak in at least two spectra of the triplicate, a specificity of 100% was achieved. The detection of KPC enzyme from inactivated bacteria impregnated in filter paper can be used as a method to confirm the presence of KPC, which could be very significant for small laboratories with limited resources.

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A novel colorimetric-fluorescent dual-mode chemosensor (JT5) based on rhodamine B has been produced for monitoring Sn4+ in the DMSO/H2O (4:1, v/v) medium. It has high sensitivity, a low detection limit, a short response time (1 s) and high stability, and can still be maintained after two weeks with the red dual fluorescence/ colorimetric response. Enhancement of red fluorescence (591 nm) and red colorimetric (567 nm) response of JT5 by Sn4+ addition. The electrostatic potential of the sensor JT5 molecule was simulated to speculate on the sensing mechanism, and the IR, mass spectrometry and 1H NMR titration were utilized to further demonstrate that JT5 was coordinated to Sn4+ with a 1:1 type, the rhodamine spironolactam ring of JT5 opens up to form a penta-membered ring with Sn4+, meanwhile, its system may have chelation enhanced fluorescence (CHEF) effect. In addition, theoretical calculations were carried out to give the energy gaps of JT5 and [JT5 + Sn4+] as well as to simulate the electronic properties of the maximal absorption peaks. Notably, the sensor JT5 was successfully applied to monitoring Sn4+ in zebrafish, and the JT5-loaded filter paper provided a solid-state platform for detecting Sn4+ by both naked eye and fluorescent methods. In summary, this work contributes to monitoring Sn4+ in organisms and solid-state materials and promotes understanding of Sn4+ functions in biological systems, environments, and solid-state materials.

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As air pollution escalates, the need for air filters increases. It is better that the filters used be based on natural fibers, such as non-wood fibers, which cause low damage to the environment. However, the short fiber lengths, low apparent densities, and high volumes of non-wood materials can make it challenging to prepare filter paper with the required mechanical and physical properties. In that context, this study focused on utilizing bamboo fibers to fabricate filter paper by employing the anthraquinone soda pulping method. The pulp underwent bleaching and oxidation processes, with the incorporation of cationic starch (CS) and polyvinyl alcohol (PVA) to enhance resistance properties, resulting in the creation of handmade filter papers. The findings revealed that the tear, burst, and tensile strength of filter paper increased with the oxidation and addition of CS and PVA. Air permeability increased with addition of PVA and combination of CS and PVA. FTIR demonstrated the conversion of hydroxyl groups in cellulose chains to carboxyl groups due to oxidation. SEM images illustrated alterations in the fiber structure post-oxidation treatment, with CS reducing pores while PVA and the CS-PVA combination enlarged pore size and enhanced porosity. The BET surface area surface area expanded with oxidation and the addition of the CS-PVA blend, indicating heightened filter paper porosity. Notably, the combined inclusion of CS and PVA not only augmented mechanical strength but also increased porosity while maintaining pore size.

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Oviductus Ranae is the dried oviduct from Rana dybowskii, a forest frog species with medicinal, tonic, and cosmetic properties. Due to the high price and resource shortage, counterfeit varieties of Oviductus Ranae often appear in the market. However, traditional identification methods cannot accurately differentiate between Oviductus Ranae and its adulterants. In this study, a rapid molecular identification method has been established. The method involves extracting genomic DNA in just 30 s using filter paper purification, species-specific rapid polymerase chain reaction (PCR) amplification, and finally, fluorescence detection of the products. It can accurately identify Oviductus Ranae and its three common adulterants in about 30 min, making the process simple, fast, and highly specific.

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Background: Malaria is a parasitic disease that affects many of the poorest economies, resulting in approximately 241 million clinical episodes and 627,000 deaths annually. Piperaquine, when administered with dihydroartemisinin, is an effective drug against the disease. Drug concentration measurements taken on day 7 after treatment initiation have been shown to be a good predictor of therapeutic success with piperaquine. A simple capillary blood collection technique, where blood is dried onto filter paper, is especially suitable for drug studies in remote areas or resource-limited settings or when taking samples from children, toddlers, and infants. Methods: Three 3.2 mm discs were punched out from a dried blood spot (DBS) and then extracted in a 96-well plate using solid phase extraction on a fully automated liquid handling system. The analysis was performed using LC-MS/MS with a calibration range of 3 - 1000 ng/mL. Results: The recovery rate was approximately 54-72 %, and the relative standard deviation was below 9 % for low, middle and high quality control levels. The LC-MS/MS quantification limit of 3 ng/mL is sensitive enough to detect piperaquine for up to 4-8 weeks after drug administration, which is crucial when evaluating recrudescence and drug resistance development. While different hematocrit levels can affect DBS drug measurements, the effect was minimal for piperaquine. Conclusion: A sensitive LC-MS/MS method, in combination with fully automated extraction in a 96-well plate format, was developed and validated for the quantification of piperaquine in DBS. The assay was implemented in a bioanalytical laboratory for processing large-scale clinical trial samples.

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AIMS: Information on breastfeeding and safety of biologics in infants is lacking due to difficulties in case collection. We evaluated methods for determining the concentration of biologics in breast milk using a dry filter method that can simplify the collection, storage and transport of breast milk. METHODS: To generate dried filter paper (DFP) samples, approximately 30 µL of breast milk was placed onto a Whatman 903 card and punched out. After extraction, the supernatant was measured using an enzyme-linked immunosorbent assay. Three concentrations of each drug were prepared in liquid breast milk (LBM) and DFP samples to determine their stability up to 28 days after storage at 2-8°C or -20°C for LBM and 25 ± 5°C for DFP. LBM and DFP samples were also provided by nursing mothers using biologics during lactation, and drug concentrations in both samples were compared. The agreement between the two measurement methods was confirmed by Bland-Altman analysis. RESULTS: Breast milk was provided by 12 mothers who used biologics (tocilizumab, abatacept, etanercept, golimumab, sarilumab and belimumab). The coefficients of variation for within-run and between-run precision for the six drugs were within 15% for both LBM and DFP, and accuracy was within 90%-110% of the quality controls. After 28 days, concentrations remained at more than 90%. The difference between the values obtained by each method was within the acceptable range of error (-12.1 to +16.6 ng/mL). CONCLUSIONS: A method for determining the concentration of biologics using DFP is expected to help improve pharmacotherapy for lactating women.

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Phase II metabolites play an important role in diazepam-related cases. The study aimed to assess the stability of diazepam's phase II metabolites in dried blood spots on filter paper. METHODS: A piece of filter paper was spotted with 100 µL of whole blood (added 1% sodium fluoride as needed) obtained from participant who received 5 mg diazepam orally, air dried for 2 h at room temperature, and then stored at different conditions. Whole spots were cut at 0.1 cm from the outer edge of blood spots at post-consumption time-points of prior (zero), 5, 16, 35, 61, 120 days and 1, 1.5 years. Analytes were extracted with methanol/water mixture (8:2, v/v) and determined using HPLC-MS/MS. Decomposition rules were analyzed by a statistical software "SPSS". RESULTS: Temazepam glucuronide remained stable (0.5-18.6% loss) at 20 â„ƒ and at 20 â„ƒ with 1% sodium fluoride for 16 days, while it was unstable after 5 days at 4 â„ƒ (21.1-26.2% loss) and - 20 â„ƒ (28.9 - 34.4% loss). After 35 days, temazepam glucuronide concentrations began to fluctuate significantly under all conditions, and an obvious increase (290.4-355.1%) was observed in 1.5 years. Oxazepam glucuronide was always unstable after 5 days, the percentage loss was even 100% when it was stored for 61 days and 1.5 years. CONCLUSIONS: Dried blood spots on ordinary filter paper are recommended to be stored at 20 â„ƒ or 20 â„ƒ with 1% sodium fluoride within 16 days. Samples should be analyzed immediately or stored in sterile and dry media.

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Although membrane separation technology has been widely used in the treatment of oily wastewater, the complexity and high cost of the membrane preparation, as well as its poor stability, limit its further development. In this study, via the vacuum-assisted suction filtration method, polydopamine (PDA)-coated TiO2 nanoparticles were tightly attached and embedded on both sides of laboratory filter paper (FP). The resultant FP possessed the typical wettability of high hydrophilicity in the air with the water contact angle (WCA) of 28°, superoleophilicity with the oil contact angle (OCA) close to 0°, underwater superoleophobicity with the underwater OCA greater than 150°, and superhydrophobicity under the water with the underoil WCA over 150° for five kinds of organic solvents (carbon tetrachloride, toluene, n-hexane, n-octane, and iso-octane). The separation efficiency of immiscible oil/water, oil-in-water, and water-in-oil emulsions using the modified FP is higher than 99%. After 17 cycles of emulsion separation, a high separation efficiency of 99% was still maintained for the FP, along with good chemical and mechanical stability. In addition, successful separation and purification were also realized for the oil-in-water emulsion that contained the methylene blue (MB) dye, along with the complete degradation of MB in an aqueous solution under UV irradiation.

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X-linked adrenoleukodystrophy (XALD) is the most common leukodystrophy. It has an estimated incidence of around 1/17.000, and a variable phenotype. Following the passage of Aidens Law, New York became the first state to implement a newborn screening for XALD in 2013. Since then, 38 American states, Taiwan, and the Netherlands have included XALD in their NBS program, and Japan and Italy have ongoing pilot studies. Screening for XALD allows for early, potentially lifesaving treatment of adrenal insufficiency and cerebral demyelination but is also a complex subject, due to our limited understanding of the natural history and lack of prognostic biomarkers. Screening protocols and algorithms vary between countries and states, and results and experiences gained so far are important for the future implementation of XALD NBS in other countries. In this review, we have examined the algorithms, methodologies, and outcomes used, as well as how common challenges are addressed in countries/states that have experience using NBS for XALD. We identified 14 peer-reviewed reports on NBS for XALD. All studies presented methods for detecting XALD at birth by NBS using a combination of mass spectrometry and ABCD1 gene sequencing. This has allowed for early surveillance of presymptomatic XALD patients, and the possibility for early detection and timely treatment of XALD manifestations. Obstacles to NBS for XALD include how to deal with variants of unknown significance, whether to screen females, and the ethical concerns of an NBS for a disease where we have limited understanding of natural history and phenotype/genotype correlation.

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Background: Some sputum smear microscopy protocols recommend placing filter paper over sputum smears during staining for Mycobacterium tuberculosis (TB) . We found no published evidence assessing whether this is beneficial. We aimed to evaluate the effect of filter paper on sputum smear microscopy results. Methods: Sputum samples were collected from 30 patients with confirmed pulmonary TB and 4 healthy control participants. From each sputum sample, six smears (204 smears in total) were prepared for staining with Ziehl-Neelsen (ZN), auramine or viability staining with fluorescein diacetate (FDA). Half of the slides subjected to each staining protocol were randomly selected to have Whatman grade 3 filter paper placed over the dried smears prior to stain application and removed prior to stain washing. The counts of acid-fast bacilli (AFB) and precipitates per 100 high-power microscopy fields of view, and the proportion of smear that appeared to have been washed away were recorded. Statistical analysis used a linear regression model adjusted by staining technique with a random effects term to correct for between-sample variability.   Results: The inclusion of filter paper in the staining protocol significantly decreased microscopy positivity independent of staining with ZN, auramine or FDA (p=0.01). Consistent with this finding, there were lower smear grades in slides stained using filter paper versus without (p=0.04), and filter paper use reduced AFB counts by 0.28 logarithms (95% confidence intervals, CI=0.018, 0.54, p=0.04) independent of staining technique. In all analyses, auramine was consistently more sensitive with higher AFB counts versus ZN (p=0.001), whereas FDA had lower sensitivity and lower AFB counts (p<0.0001). Filter paper use was not associated with the presence of any precipitate (p=0.5) or the probability of any smear washing away (p=0.6) during the staining process. Conclusions: Filter paper reduced the sensitivity of AFB microscopy and had no detectable beneficial effects so is not recommended.

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Storage of stools for the detection of soil-transmitted helminths (STH) remains challenging for the molecular diagnostic testing of STH infections. This study aimed to overcome this challenge by assessing the capacity of Whatman filter papers to store stools for the molecular detection of STHs. Stool samples were collected from school-aged children of soil-transmitted helminthiasis endemic areas of Cameroon and then, analysed using Kato Katz technique. For this study, 128 and 40 stool samples respectively with and without STH eggs were analysed. From each sample, 10, 20, 40 and 80 mg of stool were weighted and spread on 6 grades of Whatman filter papers that were stored at room temperature from one to ten weeks. DNA was extracted from spread stool using CTAB based-method. The amount of stool to spread on filter papers and the grade of filter paper offering good storage were determined by amplifying specific DNA fragments of Ascaris lumbricoides. The capacity of filter papers to store stool samples for several weeks before the molecular detection of STH species was assessed by amplifying specific DNA fragments of different STHs. The amplification rates of A. lumbricoides were significantly higher (P < 0.0001) for 10 and 20 mg of stored stools. Stools spread on Whatman paper grade 2 yielded the highest amplification rate of 100% for A. lumbricoides, T. trichiura and hookworm. PCR revealed STH infections in all the 128 spread stools carrying STH eggs. It also revealed Necator americanus and Ancylostoma duodenale respectively in 10 and 13 of 15 spread stools contained hookworm eggs. PCR confirmed the co-infections of these hookworm species as well as that of A. lumbricoides and Trichuris trichiura in 7 spread stools. Out of 40 stools without STH eggs, PCR revealed that 5 (12.5%) and 9 (22.5%) had respectively A. lumbricoides and T. trichiura infections. The amplification rate of each STH species was 100% from one to 8 weeks and decreased to 86.7% after 10 weeks of storage. This study highlighted the capacity of filter papers to store stools for the molecular detection of STHs. Storing stools on these papers will enable to monitor and evaluate control programs and ensure post-elimination surveillance.

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Background: Among the challenges in schistosomiasis surveillance and mapping surveys is the lack of a sensitive diagnostic method especially in low transmission setting. Currently, the WHO recommends the use point-of-care circulating cathodic antigen (Schisto POC-CCA) tests for surveillance and mapping of intestinal schistosomiasis. However, Schisto POC-CCA test has its drawbacks, one of which is the timely availability of test kits. One approach to overcoming this challenge is to develop a low-cost sampling method that allows for the collection and transport of urine specimens even in resource-limited settings. Objective: To develop a simple and efficient method for the collection and detection of Schistosoma mansoni (S. mansoni) CCA using urine spotted onto filter paper. Methodology: To develop a dried urine spot (DUS) method, various dried matrix extraction parameters were tested and optimized using predesigned steps. The parameters include the size of filter paper (determined by the number of punches), volume of solvents, and type of solvent. Moreover, we optimized the incubation conditions (time and temperature). Urine and stool specimens to conduct the experiments were collected from volunteer fishermen in Mwanza and this project staff. Data were entered into the Microsoft Excel spreadsheet and IBM Statistical Package for the Social Sciences, version 20 for analysis. Results: The optimal results were obtained when the procedure was run under the following conditions: Five punches of filter paper containing DUS were dissolved in 150 µl of distilled water and incubated at room temperature for 24 hours in an Eppendorf tube. More than 93% of the assays performed under these conditions produced results that were either comparable to or significantly better than the standard method. Conclusion: This study demonstrates the feasibility of collecting urine specimen (DUS) using filter paper and detecting Schistosoma CCA from DUS specimen using the Schisto POC-CCA cassette test.

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The frequency of arthropod-borne viral disease in naïve hosts is subject to change based on complex interactions among vector, host, virus, and external factors (e.g., climate). Thus, continual monitoring for both disease susceptibility and host infection dynamics is needed, especially for viruses that have proven detrimental to the health of wildlife hosts of conservation concern. The Greater Sage-grouse (Centrocercus urophasianus) is a gamebird of ecological and economic importance in the western United States for which population declines have led to a Near Threatened conservation status. Although these declines have mainly been attributed to habitat loss, West Nile virus (WNV) also poses a threat, with regional transmission potentially fueled by anthropogenic landscape alterations that may facilitate mosquito breeding. With limited WNV monitoring in Greater Sage-grouse after recognition of high susceptibility to mortality early after its initial detection in the western United States, the potential long-term impacts of WNV on this species are poorly understood. We used the plaque reduction neutralization test of filter paper strip-eluted sera to assess for anti-WNV antibodies, indicating prior infection, in opportunistically collected samples. From 2020 to 2022, 85 Greater Sage-grouse in Wyoming were sampled; none had anti-WNV antibodies. This result corroborates findings of previous studies documenting low seroprevalence. With the tenuous conservation status of the species, all potential population health risks should be considered in future management strategies, especially in the face of changing climate and landscapes.

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Dried serum spots that are well prepared can be attractive alternatives to frozen serum samples for shelving specimens in a medical or research center's biobank and mailing freshly prepared serum to specialized laboratories. During the pre-analytical phase, complications can arise which are often challenging to identify or are entirely overlooked. These complications can lead to reproducibility issues, which can be avoided in serum protein analysis by implementing optimized storage and transfer procedures. With a method that ensures accurate loading of filter paper discs with donor or patient serum, a gap in dried serum spot preparation and subsequent serum analysis shall be filled. Pre-punched filter paper discs with a 3 mm diameter are loaded within seconds in a highly reproducible fashion (approximately 10% standard deviation) when fully submerged in 10 µl of serum, named the "Submerge and Dry" protocol. Such prepared dried serum spots can store several hundred micrograms of proteins and other serum components. Serum-borne antigens and antibodies are reproducibly released in 20 µl elution buffer in high yields (approximately 90%). Dried serum spot-stored and eluted antigens kept their epitopes and antibodies their antigen binding abilities as was assessed by SDS-PAGE, 2D gel electrophoresis-based proteomics, and Western blot analysis, suggesting pre-punched filter paper discs as handy solution for serological tests.

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