Abstract
A new plasmid vector, pCRP, allowing the expression of human recombinant monoclonal antibody Fab fragments fused with a bacterial acid phosphatase has been constructed. pCRP can accept heavy- and light-chain cDNAs cloned from combinatorial antibody libraries displayed on filamentous phages with the pCombIII system and is able to direct expression to soluble Fabs in which the carboxy-terminus of the heavy chain is fused to the amino-terminus of the mature PhoN nonspecific acid phosphatase of Providencia stuartii. Using the pCRP vector, we expressed two different human recombinant Fabs cloned from combinatorial libraries (one anti-tetenus toxoid and the other anti-HIV-1 gp120) fused with the acid phosphatase. In both cases chimeric antibodies were obtained which retained the antigen-binding ability and the enzymatic activity. Similar Fab-enzyme fusions can be successfully used, even unpurified, in enzyme immunoassays.
| Original language | English |
|---|---|
| Pages (from-to) | 127-133 |
| Number of pages | 7 |
| Journal | New Microbiologica |
| Volume | 18 |
| Publication status | Published - 1995 |
Keywords
- Acid Phosphatase
- Antibodies, Monoclonal
- Base Sequence
- Gene Library
- Genes, Bacterial
- Genetic Vectors
- HIV Envelope Protein gp120
- HIV-1
- Humans
- Immunoglobulin Fab Fragments
- Molecular Sequence Data
- Providencia
- Recombinant Fusion Proteins
- Tetanus Toxoid
