ANTIBODIES AS DIAGNOSTIC TOOLS Flashcards
what can be attached to antibodies to aide the diagnosis?
Enzymes: e.g. peroxidase, alkaline phosphatase
Fluorescent probes: dyes, beads of different sizes
Magnetic beads: e.g. purification of cell types
Drugs: e.g. Kadcyla, anti-HER2 linked to emtansine
what is indirect labelling?
primary antibody detects antigen
secondary antibody with a reporter attached attaches to the primary antibody- antiantibody
how are monoclonal antibodues generated?
a mouse is challenged with an antigen and its spleen cells are cloned with myeloma cells
this produces hybridomas which are cultured in HAT medium select for positive cells
the positive cells are cloned by limited dilution
the monoclonal antibodies are harvested
what are some uses of manufactured antibodies?
Therapeutic
Prophylactic protection against microbial infection
Anti-cancer therapy
Removal of T-cells from bone marrow grafts
Block cytokine activity
Anti-calcitonin gene-related peptide (CGRP) for migraine
Diagnostic Blood group serology Quantitative immunoassays -hormones -antibodies -antigens Immunodiagnosis -Infectious diseases -Autoimmunity -Allergy (IgE) -Malignancy (myeloma)
what is an ELISA?
Enzyme Linked ImmunoSorbent Assay.
antibodies used to bind to target antigen
once all bound everything else is washed away
then a detection antibody it added which gives a signal directly proportional to the amount of antibody bound to the antigen
how might someone be HIV positive without being directly infected by the virus?
the virus may ahve crossed the placenta if the mother was HIV positive
what is flow cytometry?
sample containing cells labelled with fluorescent antibody
the sample containing cells or particles is suspended in a fluid and injected into the flow cytometer instrument. The sample is focused to ideally flow one cell at a time through a laser beam and the light scattered is characteristic to the cells and their components
