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We describe the application of a CCD Imaging Luminometer (CCDIL) for the detection and quantitation of rapid, simultaneous, peroxidase-linked chemiluminescent immunoassays of multiple samples of human serum alphafetoprotein (AFP). Results from some analogous immunoassays (total IgE, and thyroid stimulating hormone (TSH)) are included for comparison. Values for precision and antigen concentration obtained using the CCDIL and colorimetric versions of the immunoassays, on human serum samples, were in good agreement. The flexibility of the CCDIL is demonstrated; its ability to detect and quantitate antigen (particularly AFP) on a variety of solid phases is indicated. The work on the AFP immunoassays illustrates not only the flexibility of the CCDIL for sample presentation on a variety of solid phase systems, but also some relative merits of such systems. 相似文献
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Peter Jackson 《Molecular biotechnology》1996,5(2):101-123
The glycans of glycoconjugates mediate numerous important biological processes. Their separation and structural determination
present considerable difficulties because of the small quantities that are available from biological sources and the inherent
difficulty of analyzing the wide variety of complex structures that exist. A method for the analysis of reducing saccharides
by PAGE that uses specific fluorophore labeling and is simple, rapid, sensitive, and readily available to biological researchers,
has been developed. The method is known acronimically either as PAGEFS (PAGE of Fluorophore-labeled Saccharides) or in one
commercial format as FACE (Fluorophore-Assisted Carbohydrate Electrophoresis). In the PAGEFS method, saccharides having an
aldehydic reducing end group are labeled quantitatively with a fluorophore and then separated with high resolution by PAGE.
Two fluorophores, 8-aminonaphthalene-l,3,6-trisulfonic acid (ANTS) and 2-aminoacridone (AMAC), have been used to enable the
separation of a variety of saccharide positional isomers, anomers, and epimers. Subpicomolar quantities of individual saccharides
can be detected using a sensitive imaging system. Mixtures of oligosaccharides obtained by enzymatic cleavage from glycoproteins
can be labeled and electrophoresed to yield an oligosaccharide profile of each protein. AMAC can be used to distinguish unequivocally
between acidic and neutral oligosaccharides. Methods for obtaining saccharide sequence information from purified oligosaccharides
have been developed using enzymatic degradation. Other applications and the potential of the system are described. 相似文献
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