A comparative analysis of polyurethane hydrogel for immobilization of IgG on chips.
Hydrogels are considered an optimum material for protein chip surfaces, since they provide a quasi-liquid environment which allows protein activity to be maintained and shows good spot morphology as well as excellent immobilization capacity. In the following, we present a polyurethane (PU) chip that electrostatically binds IgG. The PU surface is optimized with regard to layer thickness (approximately 200 nm), hydrogel (2%) and immobilized antibody concentration (0.5 mg mL(-1); 0.3 ng spot(-1)), pH and ionic strength of the print buffer as well as to blocking solution. Evaluation is done in a direct IgG immunoassay using the Nexterion slide H as a reference. It is shown that higher IgG loading is achieved on the PU chip than on slide H, no matter whether 1x PBS (pH 7.2), Sörensen (pH 5.8) or Nexterion buffer was used as a spotting solution. Moreover, the crossreactivity with goat IgG, human IgG and monoclonal anti-CRP spotted in Nexterion buffer was as low as < or = 0.74% (slide H: < or = 3.34%).
Derwinska
Katarzyna
K
Austrian Research Centers GmbH-ARC, Department of Bioresources, 2444 Seibersdorf, Austria.
Gheber
Levi A
LA
Preininger
Claudia
C
D000818:Animals; D002021:Buffers; D006041:Goats; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D007074:Immunoglobulin G / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D009994:Osmolar Concentration; D011140:Polyurethanes / Q000737:chemistry*; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods*; D012996:Solutions
D002021:Buffers; D007074:Immunoglobulin G; D011140:Polyurethanes; D012996:Solutions; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate