Polyclonal Antibody Production Against HaptenStructured KDN Molecule by Using Different Adjuvants Alternative to Freund’s Adjuvant

Abstract

Objective: KDN (2-keto-3-deoxy-D-glycero-D-galacto-nononic acid), a member of the sialic acid family, is a hapten-structuredlow-molecular-weight monosaccharide on the cell membrane, which cannot induce immune responses without a carrier protein.Since it is over-expressed on the cancerous cells’ membrane, it is thought to be a great target molecule for anti-cancer treatments.The aim of this study is to obtain high titersof the anti-KDN polyclonal antibody response without using any carrier protein againstthe hapten-structured KDN molecule alternative to Freund’s adjuvants.Methods: Montanide™ ISA 61 VG, a water-in-oil adjuvant; ISA 201 VG, a water-in-oil-in-water emulsion adjuvant; and IMS 1313VG NPR, an aqueous-dispersion-based nanoparticle (50-200 nm) microemulsion adjuvant; and Freund’s adjuvant were used asanti-KDN antibody response stimulators. FourBALB/c mice were used for each adjuvant group, and immunization was performedat eight different time points. Anti-KDN antibody levels induced after each immunization with different adjuvants were detectedwith indirect enzyme-linked immunosorbent assay.Results: The adjuvant efficiency of Montanide™ ISA 61 VG water in oil adjuvant was 1.4 times higher than in Freund’s adjuvant(p