Biotech Jan 2010
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What happens when the body's lymphoscytes fail to the recognize its own cells and tissues as such.
Two types of autoimmune diseases
Organ specific- Brain, Thyroid, Stomach, adrenal, pancreas
Non-Organ specific- muscle, kidney, skin, jointss
AI diseases Methodologies employed at IMMCO
- -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)
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
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.
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.
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
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)
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
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.
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
What is Hep 2 used as?
Broad screening device for connective tissue disorders. (more appropriate for developing countries)
dsDNA (Immco's competitors are?)
- Inova/Binding site
What would you like to do?
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