Immunoassay of low to moderately abundant anionic proteins utilizing selective immobilization to chitosan-coated plates.
The quantitation of low to moderately abundant serum proteins is a common problem encountered in biochemistry. A physical property of the antigen of interest needs to be exploited in the initial binding step of an immunoassay resulting in capture (purification). We describe a two-stage immunoassay utilizing chromogenic and chemiluminescent substrates which is applied to two serum anionic proteins. In this assay, anionic serum proteins are selectively bound to positively charged chitosan-coated polystyrene plates at pH 6.1, in the presence of detergent. The assay has a detection limit of 0.1 microg/mL and a 1000-fold range.
Division of Rheumatology and Arthritis Centre, Department of Medicine, Queen's University, Kingston, ON K7L3N6, Canada. reesmilton@sympatico.ca
Karen J
KJ
Rees-Milton
Tassos P
TP
Anastassiades
D001798:Blood Proteins / Q000737:chemistry* / Q000378:metabolism; D048271:Chitosan / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D007118:Immunoassay / Q000379:methods*; D013379:Substrate Specificity
D001798:Blood Proteins; D055767:Immobilized Proteins; D048271:Chitosan