Fractionation of polyclonal antibody by isoelectric focusing—Differences in cross-reactivity and affinity of rabbit clonotype anti-human thyrotropin antibody
✍ Scribed by Yuichi Endo; Kiyoshi Miyai; Naoshige Hata; Kiyoshi Ichihara
- Publisher
- Elsevier Science
- Year
- 1984
- Tongue
- English
- Weight
- 734 KB
- Volume
- 143
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
Immunoglobulin G (IgG) fractions prepared from three different batches of rabbit antihuman thyrotropin (hTSH) antisera were fractionated by agarose isoelectric focusing (IEF) in the pH ranges 3 to 10 and 5 to 8. Staining of protein in agarose gel after IEF showed that polyclonal IgG separated into more than 20 protein bands with isoelectric points (~1s) ranging from 6 to 9. The clonotype antibodies to hTSH were recovered from the fractions and subjected to radioimmunoassay for determination of the binding-affinity for hTSH and the crossreactivity with human chorionic gonadotropin (hCG). The affinity constants of the antibodies recovered ranged from 6.4 X 10 M-' to 3.1 X 10" M-', and the cross-reactivities of the clonotype antibodies differed greatly. A good correlation was observed between the pls of antibody molecules and their cross-reactivities: antibodies with higher pZs bound hCG more strongly than those with lower pls. The correlation coefficients between the pls and crossreactivities were 0.83, 0.84, and 0.87 in three batches of antibody. o 1984 Academic press, IIIC.