Hybridoma Techniques Antibodies

Introduction to Hybridoma Techniques
Antibodies
highly specific proteins - bind to "antigens" specificity is a very important property of antibodies some uses of antibodies
Antibodies and therapy New Developments in Monoclonal Antibodies Are Hopeful
differentiate between very similar molecular structures detect presence of various molecular structures diagnose and treat certain cancers neutralize biological toxins (e.g., tetanus toxin) hundreds of clinical lab tests purify mixtures of molecular substances (affinity chromotography) identify microorganisms detect infected cells selectively eliminate cells enhance phagocytosis inhibit cellular interactions interfere with virus infections
antibodies are produced by stimulated B-lymphocytes
one B-lymphocyte secretes antibodies of only one specificity
desire large amounts of a single specific antibody
poly-clonal antibodies (produced in vivo)
mixture of many different antibodies of many different specificities many different B-lymphocytes producing antibodies limited time for antibody production (animal will eventually die) low levels of antibodies (animal normally produces only what is needed by that animal)
expensive to maintain animals
dependent on existing genetics of animal
cannot easily select antibodies with different biological properties
mono-clonal antibodies
eliminate mixture of antibodies by isolating B-lymphocytes in tissue culture
grow one isolated B-lymphocyte ---->one " clone" of identical B-cells
problems encountered
isolated B-lymphocyte will often not remain viable in culture don't divide very rapidly very little antibody is produced
need to "add"
viability in tissue culture rapid cell division increased antibody production
tumor cells
many types remain very viable in tissue culture most divide very rapidly but, DON'T produce any antibodies
Combine properties of
B-lymphocyte
+
Tumor Cell
produces antibodies
tissue culture viability rapid cell division
"fuse" B-lymphocyte + tumor cell into one cell
---> “hybrid" cell
culture hybrid cell
continue to grow indefinitely in culture rapid cell division (produces one "clone") antibody production (by one[mono] clone) large amounts of antibodies secreted can manipulate genetics of isolated lymphocytes can select antibodies with certain biological properties much less expensive than maintaining animals