Affinity Improvement of Antibody-Avidin Fusion Proteins for Biotin.
- Author:
Mi Young CHO
;
Hae Jung KIM
;
Hyun Mi CHO
;
Seung Uon SHIN
- Publication Type:Original Article
- Keywords:
Antibody;
Avidin fusion protein
- MeSH:
Avidin;
Biotin*;
Brain;
Chickens;
Enzyme-Linked Immunosorbent Assay;
Genetic Engineering;
Immunoglobulin G;
Ligands;
Molecular Weight;
Peptides;
Serum Albumin, Bovine
- From:Korean Journal of Immunology
1998;20(4):381-388
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
To generate drug delivery vector to locales in the body, genetic engineering and expression techniques have been used to produce antibody avidin fusion proteins. Chicken avidin has been fused to mouse-human chimeric IgG3 immediately after the hinge with a flexible linker (H-Flex-Av) and at the end of CH2 (CH2-Av). Fusion heavy chains were expressed with the expected molecular weight, assembled as H2L2 forms with a co-expressed light chain, and were secreted. The expression level of H- Flex-Av was 1~10 ug/ml/10(8)/24 hrs, but that of C2-Av was a very little (0.08~0.9 ug/ ml/10(8)/24 hrs). The resulting H-Flex-Av and CH2-Av fusion proteins continued to bind antigen dansyl and also bound biotinylated bovine serum albumin; both H-Flex-Av and CH2-Av had shown to retain 3-4 times higher relative affinity than that of CH3-Av in ELISA. Importantly the fact that both avidin fusion proteins had a higher relative affinity suggests that these avidin fusion proteins can be effectively used to deliver biotinylated ligands such as drugs and peptides to a certain locale, such as the brain.
