immunology Flashcards
what is attached to section of antibody that doesn’t bind to antigen
reporter or drug
examples of reporter
enzymes, fluorescent probes, magnetic beads
examples of enzymes
peroxidase, alkaline phosphatase
examples of fluorescent probes
dyes, beads of different sizes
what are magnetic beads used for
purification of cell types
2 examples of drugs
Kadcyla, anti-HER2 linked to emtansine
what allows antibodies to target antigens for diagnostic tests
unique specificity
what can antibodies bind to
antigens, other antibodies
what are anti-antibodies
target primary antibodies of other species
when are antibodies produced by patient
in autoimmune disease, for defence against infection
3 techniques to manufacture antibodies
antisera from immunised animals (polyclonal), monoclonal antibodies, genetically engineered antibodies
disadvantage of antisera from immunised animals
limited amount produced by animal
how are monoclonal antibodies generated
immunise animal with antigen -> remove spleen cells for antibody-producing B-cells -> remove myeloma cells (continue deviding indefinitely) -> fuse B-cells and myeloma cells with detergent to produce hybridomas -> culture in HAT medium and select for positive cells -> clone by isolating each cell (limiting dilution) -> harvest monoclonal antibodies
production of antibodies using recombinant DNA technology
place in bacteriophage -> displayed on surface of bacteriophage -> diverse range, and one with specific binding will bind and others will wash off MORE (SLIDE 11)
benefit of production of antibodies using recombinant DNA technology
expand massively when growing in bacteria
therapeutic uses of manufacture antibodies
prophylactic protection vs microbial infection, anti-cancer therapy, removal of T-cells from bone marrow grafts, block cytokine activity
example of antibodies used for prophylactic protection vs microbial infection
IVIG, synagis (anti-RSV)
example of antibodies used for anti-cancer therapy
anti-HER2
example of antibodies used for removal of T-cells from bone marrow grafts
anti-CD3
example of antibodies used for blocking cytokine activity
anti-TNFa
issue of using antibodies as treatment
very expensive
diagnostic uses of manufactured antibodies
blood group serology, immunoasays, immunodiagnosis
what immunoasssays are produced using antibodies
hormones, antibodies, antigens
4 things detected by antibodies in immunodiagnosis
infectious diseases, autoimmunity, allergy (IgE), malignancy (myeloma)
what does ELISA stand for
enzyme linked immunosorbent assay
process of ELISA
add anti-antigen antibody covalently linked to enzyme -> wash away unbound antibody -> enzyme makes coloured product from added colourless substrate -> measure absorbance of light by coloured product
process of rapid testing
lateral flow assay architecture; at first strip if molecule present will bind to antibody; excess antibody will go to second strip to bind to anti-antibody to make mark to show test worked
why might serum antibodies against HIV be present without the disease
if mother had HIV, antibodies crossed placenta
immunological concern about vague aches and pains
immune complexes
immunological concern about loss of appetite and weight loss
effect of poor nutrition on bone marrow cells
immunological concern about swollen lymph nodes in neck
immune activation
immunological concern about fever, rash, small red patches (some lumpy)
acute phase, activation of complement, immune complexes
what might immune complexes cause
inflammation and complement activation; serum sickness (in circulation); glomerulonephritis (kidney); deposition at skin, joints and lungs
what might look for if immunodeficiency concern and how
serum Ig levels (serum electrophoresis, nephelometry or ELISA), specific antibodies (ELISA), lymphocyte subsets (flow cytometry)
serum electrophoresis
compare with healthy control and active immune response; if monoclonal expansion of B cells indicates malignancy so investigate if myeloma
lymphocyte flow cytometry
mixture of cells labelled with fluorescent antibody -> stream of fluid containing antibody-labelled cells -> laser to detect which antibody present on lymphocyte to know type and number (CD4+ T cell important in HIV)
natural history of HIV infection
slide 27
HIV first and second line therapy
every 3 months monitor CD4+ count and viral load, then lower by using anti-HIV drug; second line therapy if first line doesn’t work