Method of Immunoglobulin Y (IgY) Extraction for Minimally Toxic In Vivo Applications

Abstract

Although immunoglobulin (IgY) has therapeutic potential, extracts from conventional methods are generally parenterally incompatible. This present study aimed to design a method to generate an IgY composition in vivo administration. The proposed procedure incorporated the following modifications: delipidation using carrageenan, salt precipitation using sodium sulfate, desalting using gel filtration, and affinity chromatography using MgCl2 as a disrupting agent. Furthermore, specific IgY was generated against human histones (H3 or H4) or calf histones to demonstrate the maintenance of target specificity. Native polyacrylamide gel electrophoresis patterns showed increasing purity from crude to post-desalted samples. Affinity chromatography estimated specific antibodies in the crude extract at 3–8%. Using ELISA, specificity was demonstrated for plasma, serum, or bicarbonate buffer samples. No mortality was observed in Sprague-Dawley rats and BALB/c mice using OECD up-and-down limit protocol adjusted to wrote. LD50 was estimated at more than 500 mg/kg intraperitoneally and 300 mg/kg intravenously. Intravenously injected with limited doses, intraperitoneally in mice, and daily sublethal doses for up to 36 d. Curiously, nonspecific IgY – but not specific IgY – had a higher liver injury score than solvent alone for acute injections (P < 0.05). Thus, the proposed method of IgY isolation may offer a means for an IgY composition that can be compatible with both in vivo therapeutics.