A new competitive assay for monoclonal antibodies against phi X174 single stranded DNA. 1985

K Yoshikawa, and M Fukada

A new method using L-lysine-conjugated Sepharose 4B as a matrix for coupling competitors is described for analysis of the specificity of monoclonal antibodies against single stranded DNA. In this assay, competitor-coupled Sepharose 4B is first incubated with an antibody, and then the competitor is removed (together with the antibody that is bound to it) by low-speed centrifugation of the reaction mixture. The supernatant containing only unreacted antibody is then analysed by an enzyme-linked immunosorbent assay. The method is simple and does not use radioactive materials. Moreover it is not subject to the data fluctuation that is caused by the binding of competitors to microplates in a commonly used enzyme-linked immunosorbent assay employing high concentrations of competitors.

UI MeSH Term Description Entries
D010584 Bacteriophage phi X 174 The type species of the genus MICROVIRUS. A prototype of the small virulent DNA coliphages, it is composed of a single strand of supercoiled circular DNA, which on infection, is converted to a double-stranded replicative form by a host enzyme. Coliphage phi X 174,Enterobacteria phage phi X 174,Phage phi X 174,phi X 174 Phage,Phage phi X174
D011119 Polynucleotides BIOPOLYMERS composed of NUCLEOTIDES covalently bonded in a chain. The most common examples are DNA and RNA chains. Polynucleotide
D004277 DNA, Single-Stranded A single chain of deoxyribonucleotides that occurs in some bacteria and viruses. It usually exists as a covalently closed circle. Single-Stranded DNA,DNA, Single Stranded,Single Stranded DNA
D004279 DNA, Viral Deoxyribonucleic acid that makes up the genetic material of viruses. Viral DNA
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D000327 Adsorption The adhesion of gases, liquids, or dissolved solids onto a surface. It includes adsorptive phenomena of bacteria and viruses onto surfaces as well. ABSORPTION into the substance may follow but not necessarily. Adsorptions
D000914 Antibodies, Viral Immunoglobulins produced in response to VIRAL ANTIGENS. Viral Antibodies
D001667 Binding, Competitive The interaction of two or more substrates or ligands with the same binding site. The displacement of one by the other is used in quantitative and selective affinity measurements. Competitive Binding
D012685 Sepharose Agarose,Sepharose 4B,Sepharose C1 4B,4B, Sepharose C1,C1 4B, Sepharose

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