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US 5,856,113 AGranted
Antigen and method for measuring anti-erythrocyte antibody
AssigneeNippon Zoki Pharmaceutical Co., Ltd.
InventorsHaruo Yoshii, Yuriko Fukata
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AssigneeNippon Zoki Pharmaceutical Co., Ltd.
InventorsHaruo Yoshii, Yuriko Fukata
Estimating the term…
Fragments of cell membrane of erythrocyte are used as an antigen for insolubilization in enzyme immunoassay to provide a highly sensitive, excellent quantitative method for measuring anti-erythrocyte antibody. The cell membrane fragments of erythrocyte are the residual membrane fraction after removal of soluble cytoplasm proteins which interfere with the absorbance analysis of enzyme-linked immunosorbent assay (ELISA). The cell membrane fragments of erythrocyte which are used as an antigen may be prepared by destroying the erythrocytes to obtain a cell membrane fragment fraction and a cytoplasm protein fraction. The two fractions may be separated or fractionated by gel filtration, ultrafiltration or centrifugation to isolate the cell membrane fragment fraction. A solution of the cell membrane fragments of erythrocyte is substantially transparent and is useful as an insolubilized antigen for ELISA. The antigen is shelf stable for extended periods when stored in a cool, dark location. An antigen which consists of erythrocyte membrane fragments can be prepared by simple operations and, therefore, it is possible to easily obtain large quantities of antigen for insolubilization.
An enzyme immunoassay of anti-erythrocyte antibody, said immunoassay comprising reacting a sample with an insolubilized antigen consisting of an enzyme immunoassay plate and cell membrane fragments of erythrocytes coated thereon in the substantial absence of soluble cell cytoplasm protein, said fragments having been separated from soluble cell cytoplasm protein and then insolubilized in said plate prior to said reacting.
A method for measuring anti-erythrocyte antibody comprising subjecting anti-erythrocyte antibody to an enzyme immunoassay using an insolubilized antigen consisting of an enzyme immunoassay plate and cell membrane fragments of erythrocytes coated thereon, said antigen being obtained by:
A plate for an enzyme immunoassay of anti-erythrocyte antibody, consisting of an insolubilized antigen consisting of an enzyme immunoassay plate and membrane fragments of erythrocytes coated thereon which have been separated from soluble cell cytoplasm protein.
A plate for an enzyme immunoassay of anti-erythrocyte antibody, consisting of an insolubilized antigen consisting of an enzyme immunoassay plate and membrane fragments of erythrocytes coated thereon which have been obtained by destroying erythrocytes and removing soluble cytoplasm proteins, which interfere with absorbance analysis, from said cell membrane fragments.
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