| 1. |
- Borrebaeck, Carl A K, et al.
(författare)
-
Kinetic analysis of recombinant antibody-antigen interactions : Relation between structural domains and antigen binding
- 1992
-
Ingår i: Bio/Technology. - : Springer Science and Business Media LLC. - 0733-222X. ; 10:6, s. 697-698
-
Tidskriftsartikel (refereegranskat)abstract
- The relation between domain structures of recombinant monoclonal antibody fragments and their reaction kinetics was studied for the first time using a novel biosensor based on surface plasmon resonance technology. The association and dissociation rate constants of Fab, Fv and single domain (VH fragment) anti-lysozyme antibodies were determined and compared to the intact monoclonal antibody. Fab and Fv fragments showed similar reaction kinetics and had affinity constants of 6 X 109 M-1 and 25 X 109 M-1, respectively. The single domain antibody had significantly different reaction kinetics compared to the fragments consisting of paired heavy and light chain domains. The VH domain had both a higher dissociation and a lower association rate constant, which resulted in an affinity constant approximately 250 times lower than the Fab fragment. This rapid evaluation of antibody reaction kinetics should prove to be an important selection parameter when comparing antibody fragments for their utility in therapeutic or other applications.
|
|
| 2. |
- Larrick, James W, et al.
(författare)
-
Polymemse chain reaction using mixed primers. Cloning of human monoclonal antibody variable region genes from single hybridoma cells
- 1989
-
Ingår i: Bio/Technology. - : Springer Science and Business Media LLC. - 0733-222X. ; 7:9, s. 934-938
-
Tidskriftsartikel (refereegranskat)abstract
- We describe a general approach to rapidly obtain the DNA sequence encoding the variable region of any immunoglobulin chain using the polymerase chain reaction and a mixture of upstream primers corresponding to the leader sequence, and one downstream primer designed from the conserved nucleotide sequence of the constant region. The approach was applied to five different hybridomas producing human monoclonal antibodies and variable regions for both bold gamma and mu heavy chain and kappa and lambda light chain genes were successfully cloned. cDNA encoding variable regions could be amplified from single hybridoma cells isolated by micromanipulation. This approach will permit analysis of B cell clonal ontogeny, antibody diversity and lymphoma cell progression and heterogeneity. It will also facilitate structural and functional studies of immunoglobulins as well as the rapid construction of chimeric antibodies.
|
|
| 3. |
- Mattiasson, Bo, et al.
(författare)
-
`One-pot´ protein purification by process integration
- 1994
-
Ingår i: Bio/Technology. - : Springer Science and Business Media LLC. - 0733-222X. ; 12:11, s. 1087-1089
-
Tidskriftsartikel (refereegranskat)abstract
- Is product-specific linkage of extraction and precipitation in your downstream processing future?
|
|
