Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
  • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
  • G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
  • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
  • G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
  • G01N33/564—Immunoassay; Biospecific binding assay; Materials therefor for pre-existing immune complex or autoimmune disease, i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N2800/00—Detection or diagnosis of diseases
  • G01N2800/24—Immunology or allergic disorders

Definitions

• kits claim a low level of cross reactivity with ssDNA which should render good sensitivity to low, medium, and high levels of anti- dsDNA; however, as evidenced by the kit instructions, there is a lack of sensitivity in the sera sample which are borderline positives. As a consequence, these kits cannot be used to accurately monitor patients who are borderline positives. In fact, most kits require that low positives must be retested to assure confidence in the results, thereby increasing the cost to the patient. Development of an assay which is sensitive to all levels of anti-dsDNA can only be achieved by reducing non-specific binding by such components as ssDNA.
• compositions, methods, articles, and a diagnostic test kit for the selective adsorption, or affixing of dsDNA, Sm antigen, or Sm/RNP antigen, or any of the various antigens to the autoimmune antibodies previously listed, to a pre-coated plate for the effective detection of the specific autoimmune antibodies present in sera or plasma. It is a further objective of this invention to overcome the aforementioned sensitivity, subjectivity, stability, and shelf life disadvantages inherent in many of the previously described assays. It is, moreover, also an objective of this invention to provide in the form of a kit a novel, readily utilizable means for quantitative and qualitative detection of anti-dsDNA, anti-Sm or anti-Sm/RNP.
• Figure 5 This graph depicts a standardized curve based on arbitrary READS ⁇ units/ml. This curve was generated by using the ELISA format of this invention.
• the Center for Disease Control (CDC) ANA Human Reference Lot #82-001 #5 Serum (for Sm antigen) and the ANA Human Reference Lot #82-011 #4 Serum (for RNP antigen) was run on an immunodiffusion assay with median titer of 1 :64.
• An equal volume of reference sera #4 and #5 are combined to form the standard.
• This standard was serially diluted with normal sera and graphed. The 1:1 or neat dilution was assigned arbitrarily a value of 1400 READS units.
• Step 3 Assay for the Anti-dsDNA Affixed to the Wells: Standard enzyme-linked immunoassay techniques, previously described, are used forthis assay, although any suitable means of detection such as radioactive labeling, fluorescence, or the like can be employed.
• anti-human IgG and anti-human IgM induced in goats were used to ascertain whether anti-dsDNA IgG and IgM antibodies were present. Please note that other species of animal can be used to produce anti-human antibodies..
• These antisera were linked to horseradish peroxidase, an enzyme which yields a colored product whenever one of its substrates is present together with hydrogen peroxide.
• the substrate should be chosen to be consistent with the enzyme conjugated to the antibody.
• the substrate was (3,3',5,5') Tetramethylbenzidine and hydrogen peroxide.
• TMB Substrate Solution A - contains 3,3',5,5' Tetramethylbenzidine.
• Casein blocker solution (pH 7.3) was dispensed in 200 microliter increments into each well.
• the casein blocker solution consists of 2 ml glycerol, 10 g sucrose, 15 mg casein and sufficient TEN buffer to bring the volume to 100 ml.

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• Urology & Nephrology (AREA)
• Biotechnology (AREA)
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• Microbiology (AREA)
• General Health & Medical Sciences (AREA)
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• Cell Biology (AREA)
• Organic Chemistry (AREA)
• Food Science & Technology (AREA)
• Medicinal Chemistry (AREA)
• General Physics & Mathematics (AREA)
• Pathology (AREA)
• Wood Science & Technology (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
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• Biophysics (AREA)
• Tropical Medicine & Parasitology (AREA)
• Bioinformatics & Cheminformatics (AREA)
• General Engineering & Computer Science (AREA)
• Genetics & Genomics (AREA)
• Rehabilitation Therapy (AREA)
• Rheumatology (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

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• 1990
  • 1990-02-26 AU AU51868/90A patent/AU5186890A/en not_active Abandoned
  • 1990-02-26 WO PCT/US1990/001029 patent/WO1990010229A1/en not_active Ceased
  • 1990-02-26 EP EP19900904577 patent/EP0460102A4/en not_active Withdrawn
  • 1990-02-26 CA CA002050340A patent/CA2050340A1/en not_active Abandoned

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