Anti-LTNF for in vitro assay of biological toxins

Abstract

Lethal Toxin Neutralizing Factor has been isolated in purity from opossum serum by high pressure liquid chromatography (HPLC) fractionation. The amino acid sequence from the N-terminal for the first fifteen amino acids of LTNF-n is: Leu Lys Ala Met Asp Pro Thr Pro Pro Leu Trp Ile Lys Thr Glu. Antibodies to LTNF-n and synthetic peptides consisting of fifteen, ten and five amino acids from the N-terminal of the above sequence, designated as LTNF-15, LTNF-10 and LTNF-5 were produced by immunizing Balb/C mice to produce Anti-LTNF-n, Anti-LTNF-15, Anti-LTNF-10 and Anti-LTNF-5. The anti LTNF-n, anti-LTNF-15, anti-LTNF-10 and anti-LTNF-5 react immunologically with all types of toxins derived from animal, plant and bacteria and can be assayed by immunological in vitro test such as ELISA tests. Anti-LTNFs react roughly proportional to lethal dose of biological toxins under in vitro immunological ELISA test similar to the mouse bioassay test. This property of anti-LTNFs can be utilized to replace the use of animals for bioassay of toxins from animal, plant and bacteria. Anti-LTNFs can be polyclonal raised in animals or monoclonal made in hybridomas. Anti-LTNFs can be used in crude form for immunological in vitro testing. However, purified IgG from the anti LTNFs is most desirable for consistent results from the in vitro tests. In general, the in vitro use of Anti-LTNFs is a replacement for animal use as is currently required for the assay of biological toxins.

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