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Development of the screen-printed multi-biosensor for amperometric determination of LDH, ALT and AST levels in biological fluids = 스크린 프린팅 기술을 이용한 LDH, ALT 및 AST 측정용 후막형 전기화학식 멀티 바이오센서의 개발
서명 / 저자 Development of the screen-printed multi-biosensor for amperometric determination of LDH, ALT and AST levels in biological fluids = 스크린 프린팅 기술을 이용한 LDH, ALT 및 AST 측정용 후막형 전기화학식 멀티 바이오센서의 개발 / Je-Young Chang.
발행사항 [대전 : 한국과학기술원, 2000].
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등록번호

8010706

소장위치/청구기호

학술문화관(문화관)B1층 보존서고

MBS 00023

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The screen-printed multi-biosensors for amperometric determination of LDH, ALT and AST levels in biological fluids have been constructed on the basis of $NAD^+$/NADH-dependent dehydrogenase reaction. The printing ink used for the fabrication of working electrode where enzyme-catalyzed reaction takes place is composed of reagents for enzyme reaction, polymer binder, 3,4-dihydroxybenzaldehyde (3,4-DHB) as electron-transfer mediator and graphite particle as conducting material. The printing-step is followed by electrodeposition of 3,4-DHB by potential cycling from -200 mV to +300 mV vs. standard Ag/AgCl reference electrode. Voltammetric and amperometric studies show that the multi-biosensor modified with 3,4-DHB could efficiently catalyze electrochemical oxidation of NADH generated by enzyme reaction, which indicate that electrode fouling due to direct electrochemical oxidation of NADH can be bypassed through this approach. Optimal conditions for the electrochemical determination of LDH, ALT and AST have been also investigated in terms of sensitivity. The calibration curve of the multi-biosensor for LDH level exhibits linear range up to 500 U/L and the detection limit is 50 U/L. In cases of the electrochemical determination of ALT and AST levels, both calibration curves of the multi- biosensor show linear range up to 600 U/L and the detection limits are 10 U/L in common.

바이오센서란 효소-기질, 항원-항체, 세포, 수용체 등의 생물물질을 적합한 transducer에 고정화하여 대상물질을 측정하는 장치를 일컫는다. 본 연구에서는 screen-printing technology를 이용하여, 임상의학적으로 가치를 지니는 효소인 lactate dehydrogenase (LDH), alanine aminotransferase (ALT) 및 aspartate aminotransferase(AST)의 혈중농도를 측정하기 위한 전기화학식 멀티 바이오센서를 개발하였다. 개발된 바이오센서는 $NAD^+$/NADH-dependent dehydrogenase system을 적용하였으며, NADH의 직접적인 산화로 인한 문제점을 극복하고자 electron-transfer mediator로서 3,4-dihydroxybenzaldehyde (3,4-DHB)를 채용하였다. 전기화학적인 연구를 통해 3,4-DHB는 바이오센서 내에 potential cycling을 이용한 electrodeposition을 통해 고정화를 유도할 수 있음이 확인되었고, 고정된 3,4-DHB는 효소반응 결과 생성된 NADH의 전기화학적 산화를 효과적으로 촉매함을 관찰할 수 있었다. 밝혀진 사실들을 토대로 LDH,ALT 및, AST를 측정하기 위한 측정조건의 최적화에 관한 실험을 수행하였으며, 실험결과를 바탕으로 멀티 바이오센서를 이용하여 LDH, ALT 및 AST 측정실험을 수행한 결과 LDH 측정의 경우, detection limit은 50 U/L, linear range는 약 500 U/L까지였음을 관찰할 수 있었고, ALT와 AST 측정의 경우, detection limit은 10 U/L, linaear range는 약 600 U/L까지였음이 공통적으로 확인되었다.

서지기타정보

서지기타정보
청구기호 {MBS 00023
형태사항 v, 54 p. : 삽화 ; 26 cm
언어 영어
일반주기 저자명의 한글표기 : 장제영
지도교수의 영문표기 : Hak-Sung Kim
지도교수의 한글표기 : 김학성
학위논문 학위논문(석사) - 한국과학기술원 : 생물과학과,
서지주기 Reference : p. 52-54
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Schematic representation of a generalized biosensor.

Schematic representation of screen-printing,

A set of screen-printed multi-biosensors constructed in one batch process.

Designed patterns of screen-stencils for A. Ag conducting path, B. base carbon electrode, C. pseudoreference electrode, D. insulating layer, E. working electrode.

Fabrication process of the multi-biosensor.

Proposed reaction scheme of the thick-film LDH biosensor.

Proposed reaction scheme of the thick-film ALT (AST) biosensor.

Cyclic voltammograms of the 3,4-DHB-nodified biosensor. A. Immobilization process of3,4-DHB via electrodeposition bj potential cycling between -200 mV and +300 mV ar 20 mVs. B. Cyclic voltammogram of the modified biosensor (a)before and (b)after electrodeposition of3.4-DHB (scan rate: 5mVis)

A. Cyclic voltammograms of electrodeposited 3,4-DHB at the multi-biosensor with various scan rates. (10 mV/s, 20 mV/s, 50 mVis, 100 mVls, 200 mV/s) B. Dependence of anodic peak current on scan rate at the multi-biosensor modified with 3.4-DHB

Voltammetric behavior at 10 mVls ofthe multi-biosensor modified with 3,4-DHB in the absence (a)of NADH, in the presence of (b)0.5 mM NADH, (c)1 mM NADH.

3.6 A. Voltammetric responses at 10 mVls ofthe LDH biosensor in the absence (a) orin the presence of(b)0.2 mg/mL of LDH and (c)0.4 mg/mL ofLDH. B. Voltammetric responses at 10 mVls of the GDH biosensor in the absence (al or in the nresence of 16320 mM glutamnto

A. Amperometric current responses of the LDH biosensorfor (a) 50 U/L and (b) 100 U/L ofLDH (at +300 mV). B. Amperometric current responses of the GDH electrode for (a) 100 uM and (b) 200 uM of alutamate (at +300 mV).

Dependence of the multi-biosensor response on applied

Calibration curve of the GDH electrode for glutamat (n=3, at +300 mV). Inset: linear detection range.

Dependency of thick-film GDH electrode on A. conce of a-KG and B. alanine (or aspartate) in the presence 100 mM alanine or aspartate, B:1.5 mM a-KG (n=3. a1

Effects ofincubation time on amperometric response the GDH electrode in the presence of 100 mM alanin or aspartate with 1.5 mM a-KG (n=3, at +300 mV).

Calibration curve of the multi-biosensor for LDH The inset indicates calibration curve obtained at linear detection range (n=3, at +300 mV).

Calibration curve of the multi-biosensor for ALT. The inset indicates calibration curve obtained at linear detection range (n=3, at +300 mV).

Calibration curve of the multi-biosensor for AST. The inset indicates calibration curve obtained at linear detection range (n=3, at +300 mV).

Designed patterns of screen-stencils for A. Ag conducting path, B. base carbon electrode, C. pseudoreference electrode, D. insulating layer, E. working electrode.