Abstract
A single-chain anti-atrazine antibody fragment, scAb (single-chain Fv with a CK domain), was expressed in Escherichia coli, and monomeric and dimeric species were preferentially purified from periplasmic extracts by chromatography upon nickel chelate immunosorbent columns or by immunoaffinity purification using a constant domain (CK) tag. Recombinant monomeric and dimeric antibody fragments, Fab, and intact monoclonal antibodies were compared in assays by competition between free atrazine in solution and (a) immobilized atrazine-bovine serum albumin conjugate (indirect assay) or (b) atrazine-alkaline phosphatase (direct assay). Recombinant antibody fragments provided a lower detection Limit than either Fab or intact monoclonal antibody in both assay formats. Monomeric fragments displayed a sensitivity of detection down to 0.1 ppb, compared to 1.0 ppb for dimeric fragments and the parental monoclonal.
| Original language | English |
|---|---|
| Pages (from-to) | 340-345 |
| Number of pages | 6 |
| Journal | Journal of Agricultural and Food Chemistry |
| Volume | 47 |
| Issue number | 1 |
| Early online date | 20 Nov 1998 |
| DOIs | |
| Publication status | Published - 1999 |
Keywords
- antibody fragments
- immunoassays
- single-chain antibodies
- anti-haptens
- triazines
- enzyme-immunoassay
- monoclonal-antibodies
- triazine herbicides
- atrazine
- expression
- cloning
- ELISA
- FV
- construction
- technology