| Autor: |
del Rosario RB; Department of Chemistry, University of Michigan, Medical Center, Ann Arbor 48109-0028., Baron LA, Lawton RG, Wahl RL |
| Jazyk: |
angličtina |
| Zdroj: |
International journal of radiation applications and instrumentation. Part B, Nuclear medicine and biology [Int J Rad Appl Instrum B] 1992 Apr; Vol. 19 (3), pp. 417-21. |
| DOI: |
10.1016/0883-2897(92)90128-l |
| Abstrakt: |
Disulfide links of the IgG2ak anti-ovarian carcinoma antibody, 5G6.4, were site-specifically biotinylated [approximately 2 biotins/IgG2a] using a novel crosslinking procedure using the biotin derivatized ETAC (equilibrium transfer alkylation crosslink reagent) 1a. Complexation of ETAC 1a biotinylated 5G6.4 on a column of immobilized protein A at high dilution, followed by passage of [125I]streptavidin, washing and pH change leads to elution of a streptavidin-free product with a molecular mass in the 200-300 kDa range. By contrast, direct mixing with [125I]streptavidin rapidly gave larger oligomers of much greater than 669 and approximately 440-669 kDa molecular mass, respectively. The biodistribution of the 200-300 kDa complex showed significantly diminished liver, kidney and spleen uptake as well as higher blood activity than the 440-669 kDa complex. The methodology represent the first application of ETAC chemistry to disulfide-bond directed biotinylation of antibodies and the synthesis of streptavidin antibody conjugates which minimizes their polymerization. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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