Development Of Antibody Immobilisation Strategy Using Recombinant Human Neonatal Fc Receptor
Loading...
Date
2017-08
Authors
Ng, Woei Kean
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
Antibody immobilisation is the cornerstone of antibody-based assay in biosensing technology which is predominantly used to detect and measure a wide variety of biological analytes, ranging from antigens, biomarkers, allergens and drug substances. As such, specific interactions between antibody and ligands are therefore needed to guarantee highest reproducibility and sensitivity possible. The complexity of pinning antibodies forms the key factor in interfering the outcome of an assay as the recognition molecule, i.e. the antibody itself, may have undergone structural changes and distortion when it is attached to the solid phase. In view of this, there is a need to identify new methods and novel strategy in immobilising antibody to secure the biosensing system. The present study sought to discover a new candidate of antibody binding protein, the human neonatal Fc receptor. This protein has shown its high affinity specifically towards Fc region of antibody either in vivo or in vitro. It is a heterodimer constructed of two subunits: p51 α-heavy chain (α-chain) and the β2-microglobulin light chain. Literatures have revealed its binding sites towards antibody and all of the residues involved are solely found in the α-chain. As a result, the α-chain alone was used for antibody immobilisation in the present study. The cDNA encoding for the α-chain of human neonatal Fc receptor (hFcRn-α) was cloned into a bacterial expression vector. Attempts had been made to express the soluble form of human neonatal Fc receptor in bacteria instead of the conventional approach which uses mammalian cells. The addition of betaine with a brief heat shock step during cultivation had successfully enhanced the solubility of recombinant protein expressed.
Description
Keywords
Development of antibody immobilisation strategy , using recombinant human neonatal Fc receptor