Reduction of severe bovine serum associated matrix effects on carboxymethylated dextran coated biosensor surfaces.
Surface plasmon resonance (SPR) based biosensor technology has been widely used in life science research for many applications. While the advantages of speed, ruggedness, versatility, sensitivity and reproducibility are often quoted, many researchers have experienced severe problem of non-specific binding (NSB) to chip surfaces when performing analysis of biological samples such as bovine serum. Using the direct measurement of the bovine protein leptin, present in bovine serum samples as a model, a unique buffering system has been developed and optimised which was able to significantly reduce the non-specific interactions of bovine serum components with the carboxymethyl dextran chip (CM5) surface on a Biacore SPR system. The developed NSB buffering system comprised of HBS-EP buffer, containing 0.5M NaCl, 0.005% CM-dextran, pH 9.0. An average NSB reduction (n=20) of 85.9% and 87.3% was found on an unmodified CM5 surface and a CM5 with bovine leptin immobilised on the chip surface, respectively. A reduction in NSB of up to 94% was observed on both surfaces. The concentration of the constitutive components and pH of the buffer were crucial in achieving this outcome.
Situ
Chen
C
Institute of Agri-Food and Land Use, Queen's University Belfast, David Keir Building, Stranmillis Road, Belfast BT9 5AG, United Kingdom. c.situ@qub.ac.uk
Wylie
Alastair R G
AR
Douglas
Alastair
A
Elliott
Christopher T
CT
D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods*; D002417:Cattle; D002623:Chemistry Techniques, Analytical / Q000379:methods; D003911:Dextrans / Q000032:analysis / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D020738:Leptin / Q000032:analysis; D011994:Recombinant Proteins / Q000737:chemistry; D015203:Reproducibility of Results; D012710:Serum Albumin, Bovine / Q000032:analysis* / Q000737:chemistry; D012756:Sheep
D003911:Dextrans; D020738:Leptin; D011994:Recombinant Proteins; D012710:Serum Albumin, Bovine; C014392:carboxymethyl dextran