L4 - up to complement Flashcards
why does monoclonal antibody experiments target mutant myeloma as one of the components?
it’s a B CELL TUMOR; B cells produce antibodies, antibodies are what you need to specifically select for to carry out the process you need.
mutant myeloma provides what aspect of monoclonal antibody stuff?
immortality
what does HAT stand for?
- what does aminopterin block?
- can B cells be involved in de novo/salvage pathways?
- can ONLY mutant myeloma cells survive on their own?
hypoxanthine, thymidine, aminopterin (blocks DE NOVO)
-req’d in producing NUCLEOTIDES (WANT mutant myeloma w/ B cell, not B/B or mutant/mutant)
B/B - no big deal. WON’T survive alone in HAT media
-M/M - NO! salvage pathway enzymes are MISSING and neither the thymidine or hypoxanthine pathways can be used.
What does 1) the B cell and 2) the mutant myeloma cell each give to the pathway to allow proliferation?
B cell - gives SPECIFICITY, AND enzymes req’d for MMs to grow
MM cell - now that it has enzymes to grow it can fuse w/ B cell (which wouldn’t survive) and IMMORTALITY to the combo pair
Name 6 applications of monoclonal antibody technology:
[IIIIPPF]
- ID phenotypic markers of cell types
- Immunodiagnosis
- Immunotherapy**
- Immunosuppression** (clinical med)
- Purification of antigens
- Purified isolation of unique markers
- functional analysis of cell-surface + secreted molecules
define: immunogen
substance that can INDUCE an immune response (like an antigen, just specific name)
define: hapten
- relationship to a carrier?
type of antigen that binds specific antibody, but CAN’T induce immune response on its own
carrier: TRIGGERS immune response when bound to a hapten
- vaccines! hapten allow you to be protected by vaccine
define: adjuvant
delay antigen elimination and keep it around LONGER to immune system has more time to react to it/increase its response
FOUR ways that complement can react w/ antibodies - remember - IgA does NOT activate complement!
1) cell lysis
2) phagocytosis of opsonized cell
3) complement-dependent leukocyte recruitment/inflamm
4) Fc receptor-mediated recruitment/inflamm
difference b/w affinity and avidity?
Which IG (antibody) has the highest avidity? Then which?
affinity: strength of binding an epitope and ONE SITE ONLY
avidity: COMBINED strength of binding antigen at ALL Fab sites in a molecule
IGM>IGA>IGG (purely based on number of binding sites)
two BEST WAYS to deliver an immunogen and trigger immune response:
IM and Sub-Q
-technically IP (peritoneal) applies here, but i not pleasant, or common
what is the zone of equivalence? which type of rxn is it found (precip/agglu/RIA/ELISA/FACS)?
ZofE: where immune COMPLEXES form and fall out of soln (too big now that multiple antigens/antibodies have bound)
- PRECIPITATION rxn
- forces b/w antibody-antigen are NON-COVALENT
five main fx of complement [SHILO]
OUL (earlier) still apply.
- *Solubilize immune complexes
- Humoral immunity
- promote Inflammation - C5a
- Lyse microorganisms
- Opsonization - C3b
Classical Pathway (activating complement)
-activator: ____
-binds C__
-allows opsonization of C__ to C__
(CLEAVAGE)
-then converts C__ to C__, forming the membrane attack complex (non-enzymatically) to lyse foreign bacteria
-ANTIBODY (Fc portion)–> binds C1–> converts C3 to C3b–> allows conversion of C5 to C5b –> forms membrane attack complex C5-9 to lyse antigen
Alternative Pathway (activating complement)
- activator: p___ on the m___
- DIRECTLY binds C__ in the presence of factors __ and __
- C5-C5b–> C5-C9 –> lysis
-POLYSACCHARIDE on surface of MICROBE - directly binds C3b in presence of Factors D and B
