●NineH-chains=nineIgClassessubclasses.pptVIP

* Immunology 2008 Lecture 8 Immunoglobulin Genetics 14 October ● Nine H-chains = nine Ig Classes/subclasses ● Two L-chain “types”, k l ● Four l-chain isotypes ● Allotypic variants for k and some H-chains IDIOTYPES variation unique to the Ab combining site ISOTYPES variants present in all individuals. k vs. l g1 vs. g2 [b-Hb vs. a-Hb] ALLOTYPES heritable (allelic) variants between individuals Km(1) vs. Km(2) G1m(1) vs. G1m(2) [b-Hbwt vs. b- HbS] Variation among Immunoglobulins MONOSPECIFIC Having specificity for a “single” target ? rabbit anti-kappa is monospecific relative to an antiserum to whole Ig ? rabbit anti-HSA(absBSA) is monospecific relative to the unabsorbed antiserum Neither of these is monoclonal MONOCLONAL Structurally homogeneous ● myeloma protein, Bence-Jones protein ● hybridoma Ab Isotype: Detecting and quantitating Ig in serum in healthy or diseased subjects (How much IgG or IgM…?) Allotype: Genetic studies - determining paternity, determining the source of Ig in chimeras Idiotype: Tumor immunotherapy (Ig-bearing lymphomas) Uses of anti-Isotype, -Allotype -Idiotype Analysis of light-chain Isotypes ? Start with a series of B-J proteins (monoclonal representatives of Ig light chains) ? Produce a rabbit antiserum to each one ? Test each of these antisera by Ouchterlony. Bence-Jones Protein A B C D E F G H ______________________________________________________ Rab a A + + - + - + + - Rab a B + + - + - + + - Rab a C - - + - + - - + Rab a E - - + - + - - + Rabbit Antisera Distinguish Two Kinds of B-J Proteins (A, B etc. = “kappa”; C, E etc. = “lambda”) All human Ig lights chains are either kappa or lambda. All normal human sera react with both anti-kappa and anti-lambda antisera. Therefore, kappa and lambda are isotypes. Each isotype is encoded by a unique gene… Analysis of anti-kappa antisera Absorption of antisera distinguishes two kinds of kappa chains X + Y X Y Human kappa lights chains are either Km1+ or Km1-. Normal