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USASK / OTHER / MCIM 321 / What is hybridoma?

What is hybridoma?

What is hybridoma?

Description

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"}.lst-kix_wv9gjbc57hwh-1 > li:before{content:"" counter(lst-ctn-kix_wv9gjbc57hwh-1,lower-latin) ". "}.lst-kix_wv9gjbc57hwh-2 > li:before{content:"" counter(lst-ctn-kix_wv9gjbc57hwh-2,lower-roman) ". "}.lst-kix_wv9gjbc57hwh-3 > li{counter-increment:lst-ctn-kix_wv9gjbc57hwh-3}ol.lst-kix_wv9gjbc57hwh-3.start{counter-reset:lst-ctn-kix_wv9gjbc57hwh-3 0}.lst-kix_wv9gjbc57hwh-7 > li:before{content:"" counter(lst-ctn-kix_wv9gjbc57hwh-7,lower-latin) ". "}ul.lst-kix_b47mx49hddo0-4{list-style-type:none}ul.lst-kix_b47mx49hddo0-3{list-style-type:none}ul.lst-kix_b47mx49hddo0-2{list-style-type:none}ul.lst-kix_b47mx49hddo0-1{list-style-type:none}ul.lst-kix_b47mx49hddo0-0{list-style-type:none}.lst-kix_wv9gjbc57hwh-6 > li:before{content:"" counter(lst-ctn-kix_wv9gjbc57hwh-6,decimal) ". "}.lst-kix_wv9gjbc57hwh-4 > li:before{content:"" counter(lst-ctn-kix_wv9gjbc57hwh-4,lower-latin) ". "}.lst-kix_wv9gjbc57hwh-5 > li:before{content:"" counter(lst-ctn-kix_wv9gjbc57hwh-5,lower-roman) ". 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  • Ag = antigen
  • Ab = antibody

If you want to learn more check out What happen in french revolution?

Complement:

  • Antibody bonds and complement series (amplification) bonds
  • One compliment has a hole (donut shape) and is very unstable. Thus enters the bacterial membrane (causes hole in membrane) and kills it

Antibodies are immune gamma globulin (lgG): proteins which decreased in the gel - electrophoresis gamma - globulin peak after immune serum absorption with the antigenIf you want to learn more check out How long did the black death last?

Hastens: able to bind to antibodies but don’t induce antibody formation unless coupled to an immunogenic carrier, small moleculesIf you want to learn more check out What are the active regions on chromosomes?

Chemically coupled hapten and immunogenic carrier: hapten antibody formation causing a precipitation reaction

If you want to learn more check out What is scholastic aristotelian philosophy?

Precipitation reactions are not classical chemical reactions:

  1. Bell shaped relationship between amount of precipitation and increasing antibodies in a constant antigen mix

  1. Ratios of antibody to antigen changed in the precipitates in different amounts of antibody (law of fixed proportions violated)

  • These exceptions are due to antibodies and antigens forming a lattice structure with an optimal 2 - 5 : 1 AB: AG ratio

If you want to learn more check out What is isaac newton famous for?

Evidence for lattice hypothesis:

  • Hapten - protein conjugates form precipitations only with their coupled protein and can be blocked with excess haptens that are monovalent (block antibody site)
  • lgG antibody molecules are found divalent by equilibrium dialysis
  • H1 - h2 conjugates will precipitate with both antiserum of h1 and antiserum of h2 (all three), but just h1 or just h2 will not precipitate with h1 - h2 conjugate
  • Synthesized divalent haptens form small Δ or 🗖 lattices
  • Some anti - serum will only precipitate with antigens if diluted size of the antibody antigen combining site:
  • Required for universality and specificity
  • Found to be complementary to 5 - 6 glucose in dextron polyglucose antibody

We also discuss several other topics like How do animals help us?

Hybridomas: used to get large amounts of specific antibodies by combining / fusing myelomas with spleen cells

  • Amount of antibody excess is related to the size of antigen (more precipitation based on antigen size)
  • All antibodies consist of 2 light chains and 2 heavy chains held together by disulfide bonds
  • Heavy chain can be cut by papain or pepsin at specific location
  • 2 classes of light chains = k + ⋋ (different constant regions)
  • 7 classes of heavy chains = lgM, lgE, lgAm lgG 1-4

  • Cells make either k or ⋋ light chains (specialized)
  • lgM + lgA have J chain (function still unknown)
  • Immuno absorbent column are used to isolate antibodies with specific heavy chains

Treating foreign antibodies as antigens, lgA was found to have slight differences (a - chain in constant region) which are only 1 - 2 epitopes and won’t cause antibody formation in different species, but will in the same species, called allotypes: antibody which recognizes specific differences from the genome of the same species

  • These allotype epitopes are inherited using simple Mendelian laws (one gene controls them)
  • Determinant are different in all lgA molecules vs lgG molecules vs ⋋ vs k
  • Cell can only make one or another marker on heavy chains (simple Mendelian inheritance) in inbred strains
  • You receives a genetic “lego set” from your parents (to assemble antibodies) made of parts / fragments of genes (for the variable region which is attached to an inherited constant region with allotypes) which the cell can select to code for antibodies
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