Engineered recombinant single-chain fragment variable antibody for immunosensors

Zhihong Shen, Raymond L. Mernaugh, Heping Yan, Lei Yu, Ying Zhang, Xiangqun Zeng

Research output: Contribution to journalArticlepeer-review

35 Scopus citations

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 × 10-9-3.3 × 10-8 M. The piezoimmunosensor was also used to study the kinetics and thermodynamics of antigen/scFv antibody binding.

Original languageEnglish (US)
Pages (from-to)6834-6842
Number of pages9
JournalAnalytical Chemistry
Volume77
Issue number21
DOIs
StatePublished - Nov 1 2005
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

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