Biotech Jan 2010

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Author:
Doug
ID:
5053
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Biotech Jan 2010
Updated:
2010-01-25 00:08:55
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Biotech
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Description:
Biotech Q/A drawn from Suresh's presentation
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  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

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