Preparation of monospecific immunoglobulin against human leukocytic interferon

Largomycin has been purified to homogeneity by chromatography on hydroxylapatite whereby carbohydrate and protease impurities were removed. Largomycin is an acidic protein (pI 4.13, molecular weight 29300) which forms a dimer in phosphate buffer. An N-terminal amino acid sequence analysis from the amino-terminal residue gave, for the first 32 residues, Asp-Ile-Leu-Ile-Ala-Gly-Ala-Thr-Gly-Asn-Val-Gly-Lys-Pro-Leu-Val-Glu-Gly-Leu-Leu - Ala-Ala-Gly-Lys-Pro-Val-Arg-Ala-Leu-Thr-Arg-Asn... The sequence from the carboxyl terminus was -Ala-Ala-Leu-Phe-OH with threonine, valine, and glutamic acid being released upon prolonged digestion. The same amino acid sequences were found for largomycin prepared from either the culture broth or the mycelium of Streptomyces pluricolorescens. The similarities extended to the other physical properties, the antimicrobial activity against Staphylococcus aureus and Sarcina lutea, and the antitumor activity against KB cells. Largomycin inhibits the biosynthesis of DNA and RNA. An iodinated derivative did not bind to KB cells. The antimicrobial activity was lost following ultraviolet irradiation, protection against which was not afforded by p-aminobenzoic acid.

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“…Immunopotentiation Assay. Immunopotentiation activity was measured as described by Skurkovich et al (1978) and Herberman (1977).…”

Section: Absorbancementioning

confidence: 99%