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Molecular characterization of a single-chain antibody variable fragment (scFv) specific for PspA from Streptococcus pneumoniae.
Jang, ShinA; Kim, Gyuhee; Oh, Jihye; Lee, Seungyeop; Kim, Dongho; Kim, Kook-Han; Kim, Yong Ho; Rhee, Dong-Kwon; Lee, Sangho.
Afiliación
  • Jang S; Department of Biological Sciences, Sungkyunkwan University, Suwon, 16419, South Korea.
  • Kim G; Department of Biological Sciences, Sungkyunkwan University, Suwon, 16419, South Korea.
  • Oh J; Department of Biological Sciences, Sungkyunkwan University, Suwon, 16419, South Korea.
  • Lee S; School of Pharmacy, Sungkyunkwan University, Suwon, 16419, South Korea.
  • Kim D; Department of Biological Sciences, Sungkyunkwan University, Suwon, 16419, South Korea.
  • Kim KH; SKKU Advanced Institute of Nanotechnology (SAINT), Sungkyunkwan University, Suwon, 16419, South Korea.
  • Kim YH; SKKU Advanced Institute of Nanotechnology (SAINT), Sungkyunkwan University, Suwon, 16419, South Korea.
  • Rhee DK; School of Pharmacy, Sungkyunkwan University, Suwon, 16419, South Korea.
  • Lee S; Department of Biological Sciences, Sungkyunkwan University, Suwon, 16419, South Korea. Electronic address: sangholee@skku.edu.
Biochem Biophys Res Commun ; 482(1): 141-146, 2017 Jan 01.
Article en En | MEDLINE | ID: mdl-27845043
Streptococcus pneumoniae is a major infectious agent responsible for pneumonia, otitis media, sepsis and meningitis. Pneumococcal surface protein A (PspA) is a well-characterized virulence factor localized on the surface and a target for vaccine development. In this study, we screened a single-chain antibody variable fragment (scFv) using phage display from a human synthetic library to select a clone 2B11. Affinity (Kd) of 2B11 was measured to be 5 nM using biolayer interferometry. 2B11 exhibited a dose-dependent recognition of recombinant PspA with no cross-reactivity towards pneumococcal antigens. The epitope on PspA was defined to residues 231-242 by mutational analysis. Molecular docking analysis supported the experimentally determined epitope, suggesting that the helix spanning residues 231-242 can bind to 2B11 with residues in the CDR-H3 (complementarity determining region 3 in the heavy chain) actively participating in the molecular contacts. Comparison of 2B11 with a commercial PspA antibody revealed that 2B11 exhibited a better specificity towards recombinant PspA antigen. 2B11 was capable of detecting endogenous PspA from pneumococcal lysates with affinity similar to that of the commercial antibody. Our study provides a molecular tool for biosensors detecting pneumococcal diseases.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Streptococcus pneumoniae / Proteínas Bacterianas / Anticuerpos de Cadena Única Idioma: En Revista: Biochem Biophys Res Commun Año: 2017 Tipo del documento: Article País de afiliación: Corea del Sur Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Streptococcus pneumoniae / Proteínas Bacterianas / Anticuerpos de Cadena Única Idioma: En Revista: Biochem Biophys Res Commun Año: 2017 Tipo del documento: Article País de afiliación: Corea del Sur Pais de publicación: Estados Unidos