Engineered recombinant single-chain fragment variable antibody for immunosensors.
Anal Chem
; 77(21): 6834-42, 2005 Nov 01.
Article
em En
| MEDLINE
| ID: mdl-16255580
ABSTRACT
A recombinant single-chain fragment variable (scFv) antibody (designated A10B) was engineered to contain two histidines within the linker peptide used to join the scFv heavy and light chains. A piezoimmunosensor using the scFv was successfully developed. A10B scFv bound to the gold piezoimmunosensor surface were correctly oriented, retained antigen-binding activity, and coupled at high surface concentration. These results, and results obtained from an earlier study using an scFv containing a linker cysteine, suggest that the location on the linker sequence in which the amino acids were incorporated was well tolerated by the scFv and did not interfere with scFv antigen-binding activity. The scFv-modified QCM sensor was thoroughly characterized and used to specifically detect antigen in crude serum sample and had a sensitivity of 2.3 +/- 0.15 nM (n = 4) with a linear range over 2.3 x 10(-9) - 3.3 x 10(-(8) M. The piezoimmunosensor was also used to study the kinetics and thermodynamics of antigen/scFv antibody binding.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteínas Recombinantes
/
Fragmentos de Imunoglobulinas
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Técnicas Biossensoriais
/
Engenharia de Proteínas
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Ano de publicação:
2005
Tipo de documento:
Article