Biotech Jan 2010

  1. What happens when the body's lymphoscytes fail to the recognize its own cells and tissues as such.
    Autoimmune Disease
  2. Two types of autoimmune diseases
    Organ specific- Brain, Thyroid, Stomach, adrenal, pancreas

    Non-Organ specific- muscle, kidney, skin, jointss
  3. AI diseases Methodologies employed at IMMCO
    Immunofluorescence (IF)

    • -Direct Immunofluorescence (DIF) take biopsy and put on slide
    • -Indirect Immunofluorescence (IFA) instead of my own tissue it would be mouse etc.
    • Enzyme Linked Immunosorbent Assay (ELISA)
    • Western Blot (WB)
  4. Summarize Diagnosis of Autoimmune Diseases (by IFA)
    • -Allows visualization of Multiple antibodies
    • -Screening or confirmation
    • -Can easily accomodate small to medium scale test volume
    • -Requires staff capable of reading IFA reactions
  5. How does Western Bloc detection work?
    • 1. Test strips are incubated with patient serum
    • 2. Antibodiees in serum attach to antigens on strips.
    • 3. A secondary enzyme connected to reporter enzyme, attacheds to the antigen/antibody complex.
    • 4. This entire complex in the prescense of substrate will change color and mark the position of a positive reaction.
  6. Western Bloc Diagnosis offers:
    • 1) Detection of individual or multiple antibodies
    • 2) Generally a confirmatory assay.
    • 3) Can efficiently be used in small scale test volumes
    • 4) read visually
    • 5) multiple antibodies may be detected simultaneously.
    • 6) provides a detection system for hard to purify protiens.
  7. What is Hep-2?
    • -Human epithelial cells -line 2
    • -Cells originate from a human laryngeal carcinoma
    • -The cells ar grown as a monolayer on microscope slides
    • -These slides provide a sensitive substrate
  8. There are basically 4 recognizable Patterns with Hep-2
    • -Nuclear Homogeneous- SLE (antigens dsDNA, Histones)
    • -Nuclear Coarse Speckled- MCTD, SLE, Sjogrens syndrome (antigen U1-snRNP or Sm proteins)
    • -Nucleolar- 25-50% of patients with myositis-scleroderma (PM-Scl complex antigen)
    • -Cell Cycle- Crest Syndrome (antigen is kinetochore plates)
    • -Cytoplasmic- Primary biliary cirrhosis (antigen is mitochondrial autoantigen M2)
  9. What is a diagnostic Marker?
    Marker, blood: A sign of a disease or condition that can be isolated from a blood sample. For example, the monoclonal antibody D8/17 is a diagnostic sign of Pediatric Autoimmune Disorders associated with Strep
  10. dsDNA (Double Stranded DNA) is?
    • 1) a Diagnostic marker for systemic Lupus
    • 2) Autoantigen is the nucleic acid DNA (of patient) against which antibodies are produced.
    • 3) not present in healthy individuals.
  11. dsDNA detection methods?
    • 1) IFA Clift Assay
    • 2) IFA Hep 2 (screening test, meaning you have to do more testing)
    • 3) RIA Farr Method
    • 4) EIA
  12. What is Hep 2 used as?
    Broad screening device for connective tissue disorders. (more appropriate for developing countries)
  13. dsDNA (Immco's competitors are?)
    • Inova/Binding site
    • Euroimmun
Card Set
Biotech Jan 2010
Biotech Q/A drawn from Suresh's presentation