Counting of single protein molecules at interfaces and application of this technique in early-stage diagnosis

被引:34
作者
Löscher, F
Böhme, S
Martin, J
Seeger, S [1 ]
机构
[1] Univ Regensburg, Inst Analyt Chem Cheo & Biosensor, D-93040 Regensburg, Germany
[2] SL Microtest GMBH, D-07745 Jena, Germany
关键词
D O I
10.1021/ac971378r
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The fluorescence-based detection and counting of single protein molecules after specific binding to antibodies at interfaces is presented. A diode laser was used as the excitation source. The unspecific binding at the interface has been reduced to a level of only 0.1% of the maximum signal level. At present, the detection limit of this molecule counting process is in the range of 10(-17) mol/L, and the dynamic range of the signal corresponds to 7 orders of magnitude of antigen concentration, but these values are not limiting. As a preliminary application in early-stage diagnosis, we have investigated the detection of a single cardiac actin molecule in human plasma, which is of interest in myocardial infarction diagnosis.
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收藏
页码:3202 / 3205
页数:4
相关论文
共 33 条
[1]   SINGLE MOLECULES OBSERVED BY NEAR-FIELD SCANNING OPTICAL MICROSCOPY [J].
BETZIG, E ;
CHICHESTER, RJ .
SCIENCE, 1993, 262 (5138) :1422-1425
[2]   PHOTODAMAGE TO INTACT ERYTHROCYTE-MEMBRANES AT HIGH LASER INTENSITIES - METHODS OF ASSAY AND SUPPRESSION [J].
BLOOM, JA ;
WEBB, WW .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1984, 32 (06) :608-616
[3]   LIPID DIFFUSIBILITY IN THE INTACT ERYTHROCYTE-MEMBRANE [J].
BLOOM, JA ;
WEBB, WW .
BIOPHYSICAL JOURNAL, 1983, 42 (03) :295-305
[4]   USE OF DIODE-LASERS IN AN EVANESCENT-WAVE IMMUNOSENSOR FOR STATIONARY AND TIME-RESOLVED DETECTION OF ANTIGENS [J].
BOCK, D ;
GALLA, K ;
MARTIN, M ;
WOLFRUM, J ;
SEEGER, S .
SENSORS AND ACTUATORS B-CHEMICAL, 1995, 29 (1-3) :293-299
[5]   SORTING SINGLE MOLECULES - APPLICATION TO DIAGNOSTICS AND EVOLUTIONARY BIOTECHNOLOGY [J].
EIGEN, M ;
RIGLER, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (13) :5740-5747
[6]   HIGH SPECIFIC ACTIVITY CHEMI-LUMINESCENT AND FLUORESCENT MARKERS - THEIR POTENTIAL APPLICATION TO HIGH-SENSITIVITY AND MULTI-ANALYTE IMMUNOASSAYS [J].
EKINS, R ;
CHU, F ;
MICALLEF, J .
JOURNAL OF BIOLUMINESCENCE AND CHEMILUMINESCENCE, 1989, 4 (01) :59-78
[7]   IMAGING OF SINGLE FLUORESCENT MOLECULES AND INDIVIDUAL ATP TURNOVERS BY SINGLE MYOSIN MOLECULES IN AQUEOUS-SOLUTION [J].
FUNATSU, T ;
HARADA, Y ;
TOKUNAGA, M ;
SAITO, K ;
YANAGIDA, T .
NATURE, 1995, 374 (6522) :555-559
[8]   ACTIN EXPRESSION IN SMOOTH-MUSCLE CELLS OF RAT AORTIC INTIMAL THICKENING, HUMAN ATHEROMATOUS PLAQUE, AND CULTURED RAT AORTIC MEDIA [J].
GABBIANI, G ;
KOCHER, O ;
BLOOM, WS ;
VANDEKERCKHOVE, J ;
WEBER, K .
JOURNAL OF CLINICAL INVESTIGATION, 1984, 73 (01) :148-152
[9]  
Galla K, 1994, J Fluoresc, V4, P111, DOI 10.1007/BF01876665
[10]   TIME-RESOLVED FLUORESCENCE STUDIES OF LABELED NUCLEOSIDES [J].
HAN, KT ;
SAUER, M ;
SCHULZ, A ;
SEEGER, S ;
WOLFRUM, J .
BERICHTE DER BUNSEN-GESELLSCHAFT-PHYSICAL CHEMISTRY CHEMICAL PHYSICS, 1993, 97 (12) :1728-1730