원문정보
Quantitative Assay of Recombinant Hepatitis B Surface Antigen by Using Surface Plasmon Resonance Biosensor
초록
영어
We performed a basic experiment for rapid, on-line, real-time measurement of HBsAg by using a surface plasmon resonance biosensor to quantify the recognition and interaction of biomolecules. We immobilized the anti-HBsAg polyclonal antibody to the dextran layer on a CM5 chip surface which was pre-activated by N-hydroxysuccinimide for amine coupling. The binding of the HBsAg to the immobilized antibody was measured by the mass increase detected
by the change in the SPR signal. The binding characteristics between HBsAg and its antibody followed typical monolayer adsorption isotherm. When the entire immobilized antibody was interacted, there was no additional, non-specific binding observed, which suggested the biointeraction was very specific as expected and independent of the ligand density. No significant steric hindrance was observed at 17.6 nm/mm2 immobilization density. The
relationship between the HBsAg concentration in the sample solution and the antigen bound to the chip surface was linear up to ca. 40 μg/mL, which is much wider than that of the ELISA method. It appeared the antigen-antibody binding was increased as the immobilized ligand density increased, but verification is warranted. This study showed the potential of this biosensor-based method as a rapid, simple, multi-sample, on-line assay. Once properly validated, it can serve as a more powerful method for HBsAg quantification replacing the current ELISA method.
목차
서론
재료 및 방법
HBsAg 및 anti-HBsAg 항체
리간드 항체 고정화 과정
Biorecognition에 의한 HBsAg의 결합반응 분석
칩 표면의 재생을 위한 최적조건 결정
결과 및 고찰
리간드 고정화
항원-항체 결합반응 및 선형 범위의 결정
최대결합도 및 항원-항체 결합반응의 몰 비율
재생조건의 결정
결론
REFERENCES