| | |  | | | | Record from Web of Science® | | | | | | |
| HUMAN ANTI-SELF ANTIBODIES WITH HIGH SPECIFICITY FROM PHAGE DISPLAY LIBRARIES |
|
|
| Author(s): GRIFFITHS AD, MALMQVIST M, MARKS JD, BYE JM, EMBLETON MJ, MCCAFFERTY J, BAIER M, HOLLIGER KP, GORICK BD, HUGHESJONES NC, HOOGENBOOM HR, WINTER G |
| Source: EMBO JOURNAL Volume: 12 Issue: 2 Pages: 725-734 Published: FEB 1993 |
| Times Cited: 340 References: 104 |
| Abstract: Recently we demonstrated that human antibody fragments with binding activities against foreign antigens can be isolated from repertoires of rearranged V-genes derived from the mRNA of peripheral blood lymphocytes (PBLs) from unimmunized humans. The heavy and light chain V-genes were shuffled at random and cloned for display as single-chain Fv (scFv) fragments on the surface of filamentous phage, and the fragments selected by binding of the phage to antigen. Here we show that from the same phage library we can make scFv fragments encoded by both unmutated and mutated V-genes, with high specificities of binding to human self-antigens. Several of the affinity purified scFv fragments were shown to be a mixture of monomers and dimers in solution by FPLC get filtration and the binding kinetics of the dimers were determined using surface plasmon resonance (k(on) = 10(5)-10(6) M-1 s-1, k(off) = 10(-2) s-1 and K(a) = 10(7) M-1). The kinetics of association are typical of known Ab-protein interactions, but the kinetics of dissociation are relatively fast. For therapeutic application, the binding affinities of such antibodies could be improved in vitro by mutation and selection for slower dissociation kinetics. |
| Document Type: Article |
| Language: English |
Addresses:
1. MRC, MOLEC IMMUNOL UNIT, CAMBRIDGE CB2 2QH, ENGLAND
2. UNIV CAMBRIDGE, DIV TRANSFUS MED, CAMBRIDGE CB2 2PT, ENGLAND
3. CAMBRIDGE ANTIBODY TECHNOL LTD, MELBOURNE SG8 6EJ, CAMBS ENGLAND
4. MRC, MOLEC BIOL LAB, CAMBRIDGE CB2 2QH, ENGLAND |
| Publisher: OXFORD UNIV PRESS UNITED KINGDOM, WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP |
| Subject Category: Biochemistry & Molecular Biology; Cell Biology |
| IDS Number: KL717 |
| ISSN: 0261-4189 |
|
| | | | | | | | | Record from Web of Science® | | | | | | | | | |