Glow-discharge treated piezoelectric quartz crystals as immunosensors for HSA detection.
The aim of this study is to develop an immunoaffinity sensor based on piezoelectric crystals for human serum albumin (HSA) detection in aqueous media. Quartz crystals were treated with ethylene diamine (EDA) plasma in a glow-discharge apparatus in order to substitute amino groups on their surfaces. Then anti-HSA antibodies were immobilized via these amino groups by using glutaraldehyde (GA) as cross-linker. Immobilization of the antibody on the quartz crystal was examined for different pH, antibody concentration and treatment time. The optimum conditions for anti-HSA immobilization were evaluated by the measurements of the activity of the surface against HSA. The optimum values of pH, antibody concentration and treatment time were found 6.2, 0.15 mg/ml and 2 h, respectively. For detection of HSA into the solution, two methods were used. In the first (dip and dry) method, the frequency shifts were measured in air after the 1 h interaction of the anti-HSA immobilized crystals with HSA solution. In the other (direct) method, the frequency shifts were followed continuously for 60 min. while the probe was immersed in the HSA solution. An increase for the frequency shifts was observed with increasing of HSA concentration of 16-128 microg/ml. Both the immobilization and antibody-interaction conditions were found important on the extend of these specific interaction. The relations between the HSA concentrations and frequency shifts were exponential in both methods.
Department of Chemical Engineering and Bioengineering Division, Hacettepe University, 06532 Beytepe, Ankara, Turkey.
Reza
R
Saber
Selma
S
Mutlu
Erhan
E
Pişkin
D000327:Adsorption; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D000918:Antibody Specificity; D000936:Antigen-Antibody Complex / Q000032:analysis*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D003460:Crystallization; D004563:Electrochemistry; D004867:Equipment Design; D005029:Ethylenediamines / Q000737:chemistry; D005976:Glutaral / Q000737:chemistry; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods; D011791:Quartz / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012709:Serum Albumin / Q000032:analysis* / Q000276:immunology
D000906:Antibodies; D000936:Antigen-Antibody Complex; D005029:Ethylenediamines; D012709:Serum Albumin; D011791:Quartz; C031234:ethylenediamine; D005976:Glutaral