Oriented antibody immobilization to polystyrene macrocarriers for immunoassay modified with hydrazide derivatives of poly(meth)acrylic acid

Vadim V Shmanai,Tamara A Nikolayeva,Ludmila G Vinokurova,Anatoli A Litoshka

doi:10.1186/1472-6750-1-4

Vadim V Shmanai, Tamara A Nikolayeva + Show 2 more

Open Access

https://doi.org/10.1186/1472-6750-1-4

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Abstract

BackgroundHydrophobic polystyrene is the most common material for solid phase immunoassay. Proteins are immobilized on polystyrene by passive adsorption, which often causes considerable denaturation. Biological macromolecules were found to better retain their functional activity when immobilized on hydrophilic materials. Polyacrylamide is a common material for solid-phase carriers of biological macromolecules, including immunoreagents used in affinity chromatography. New macroformats for immunoassay modified with activated polyacrylamide derivatives seem to be promising.ResultsNew polymeric matrices for immunoassay in the form of 0.63-cm balls which contain hydrazide functional groups on hydrophilic polymer spacer arms at their surface shell are synthesized by modification of aldehyde-containing polystyrene balls with hydrazide derivatives of poly(meth)acrylic acid. The beads contain up to 0.31 μmol/cm2 active hydrazide groups accessible for covalent reaction with periodate-oxidized antibodies. The matrices obtained allow carrying out the oriented antibody immobilization, which increases the functional activity of immunosorbents.ConclusionsAn efficient site-directed antibody immobilization on a macrosupport is realized. The polymer hydrophilic spacer arms are the most convenient and effective tools for oriented antibody coupling with molded materials. The suggested scheme can be used for the modification of any other solid supports containing electrophilic groups reacting with hydrazides.

Highlights

Hydrophobic polystyrene is the most common material for solid phase immunoassay
Polyacrylamide is a common material for solid-phase carriers of biological macromolecules, including immunoreagents used in affinity chromatography and immunoassay [1,2]
We investigated two types of carriers, which were produced by a modification of aldehyde groups on the surface of formylated polystyrene balls: (1) with a range of low-molecular-weight hydrazide spacer arms described in our previous paper [14], and (2) with copolymers of acrylamide and methacrylic acid hydrazide

Summary

Introduction

Hydrophobic polystyrene is the most common material for solid phase immunoassay. Biological macromolecules were found to better retain their functional activity when immobilized on hydrophilic materials. Polyacrylamide is a common material for solidphase carriers of biological macromolecules, including immunoreagents used in affinity chromatography. Polyacrylamide is a common material for solid-phase carriers of biological macromolecules, including immunoreagents used in affinity chromatography and immunoassay [1,2]. Various schemes of polyacrylamide modification to introduce applicable functional groups for covalent protein immobilization were reported: hydrolysis to form carboxylic groups (the immobilization is carried out through the protein's amino groups using condensing agents), hydrazinolysis to form hy-. As for the solid phase macro carriers (plates, balls, tubes) in immunoassay, hydrophobic polystyrene is used as the most common material. Proteins are immobilized on polystyrene by passive adsorption, which often causes considerable denaturation [4]

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