Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1997-04-24
1999-10-12
Chin, Christopher L.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 71, 435 75, 435962, 435965, 436506, 436509, 436512, 436513, 436518, 436547, 436548, 530380, 5303871, 5303881, 53038815, 530829, 530861, 530862, 530863, 530864, 530865, 530866, G01N 3353
Patent
active
059653787
DESCRIPTION:
BRIEF SUMMARY
The invention concerns a composition composed of several different antibodies or/and antibody fragments which is suitable as a reagent to reduce interferences in an immunological method for the class-specific detection of antibodies from one or several of the immunoglobulin classes G, M, A, D and E.
The mammalian organism contains various classes of antibodies which are formed by the B celli of the immune system to defend against antigens. The antibody molecules are each composed of one or several sets of four polypeptide chains, two heavy chains and two light chains which are linked together via disulfide bridges.
Antibodies are generally divided into the classes G, M, A, D and E. These five immunoglobulin classes differ in their heavy chain which is denoted .gamma., .mu., .alpha., .delta. and .di-elect cons. chain. In addition there are also immunoglobulin subclasses in the case of IgG, IgA and IgM.
Antibodies of the IgG class constitute 70 to 75% of the total immunoglobulin in normal human serum (corresponding to 8 to 16 mg/ml). They are mainly formed as the secondary immune response of the organism to an infection.
Antibodies of the IgM class make up ca. 10% of the immunoglobulin present in human serum and have a pentameric structure. These antibodies appear very early after an infection so that their determination is important for the early detection of diseases.
The immunoglobulins of the IgA class form about 15 to 20% of the immunoglobulin present in human serum and are the most important secretory immunoglobulin in saliva, milk and secretions of the urogenital region.
Antibodies of the IgD class are located on the membrane of circulating B cells and it is assumed that they play a role in autoimmune diseases.
Antibodies of the IgE class only occur in a very small amount in serum, but they play an important role in a number of allergic reactions such as asthma and hay-fever.
The class-specific determination of immunoglobulins i.e. the selective determination of antibodies of one or several selected immunoglobulin classes or subclasses which are directed against a particular antigen in the presence of antibodies of other immunoglobulin classes or subclasses which are directed against the same antigen is of particular importance for the detection of particular diseases e.g. for the early diagnosis of infections, to differentiate between acute and healed infections and to make precise prognoses.
Methods for the class-specific determination of immunoglobulins are known. For this an immune component that is specific for a selected antibody class e.g. an antibody against the .mu. chain of human IgM can be coupled to a solid carrier and the antigen-specific immunoglobulin component can be detected by reaction with a directly or indirectly labelled antigen. A disadvantage of this type of method is that non-antigen-specific immunoglobulins of the respectively selected immunoglobulin class compete with the antigen-specific molecules for the solid phase antibodies. This can result in a falsification of the results dependent on this quantity ratio.
A further disadvantage of the methods of the state of the art is that a washing step is required after incubation of the sample with the immune components on the solid carrier and before reaction with the antigen-specific immune components. This is a potential source of error and considerably increases the testing time.
EP-B-0 292 810 discloses a method for the determination of antigen-specific antibodies from one of the immunoglobulin classes M, A, D and E in a sample liquid wherein an interference-eliminating reagent is added to avoid interferences by antibodies of the IgG class which is selected from anti-human IgG, aggregated human or animal IgG or a .gamma.-Fc fragment. The interference-eliminating reagent is intended to eliminate the antigen binding of the specific IgG antibodies present in the sample and to suppress the activity of rheumatoid factors.
However, the reagents according to EP-B 0 292 810 for eliminating interference have some disadvantages. Thus in
REFERENCES:
patent: 4914040 (1990-04-01), Lenz et al.
Carrel et al., Immunochemistry. 10:245-250, 1973.
Duermeyer et al., Journal of Medical Virology. 4:25-32, 1979.
McDougal et al., J. Lab. Clin. Med. 106(1):80-87, 1985.
Powell et al., The Journal of Rheumatology, An Improved Assay for IgG Rheumatoid Factor: Its Value in the Diagnosis of Rheumatoid Arthrits, 1985, pp. 427-731.
Klemt Volker
Lenz Helmut
Schlieper Dittmar
Schmitt Urban
Chin Christopher L.
Nguyen Bao-Thuy
Roche Diagnostics GmbH
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