Generating antibodies

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(Type of antibody)
Line 8: Line 8:
'''polyclonal'''
'''polyclonal'''
* faster to generate
* faster to generate
-
* less specific)
+
* less specific
 +
* cheaper than monoclonals
'''monoclonal'''
'''monoclonal'''
Line 15: Line 16:
* after initial setup no animals required; unlimited production from immortal cell line
* after initial setup no animals required; unlimited production from immortal cell line
* antigen can be impure; subsequent selection of hybridoma clones
* antigen can be impure; subsequent selection of hybridoma clones
 +
* more expensive than polyclonals
=== Source organism ===
=== Source organism ===

Revision as of 07:41, 23 April 2008

Drawing of an antibody - heavy chains red, light chains yellow.
Drawing of an antibody - heavy chains red, light chains yellow.

You are hot in pursuit of an interesting protein but there are no commercial antibodies or they are bad. It's time to think about raising your own antibodies against the protein. It will be a time and money consuming undertaking but once you developed a specific antibody or set of antibodies, you will be in a unique position to generate interesting data.

Contents

Early decisions

Type of antibody

polyclonal

  • faster to generate
  • less specific
  • cheaper than monoclonals

monoclonal

  • slower to generate
  • more specific
  • after initial setup no animals required; unlimited production from immortal cell line
  • antigen can be impure; subsequent selection of hybridoma clones
  • more expensive than polyclonals

Source organism

  • rabbit
  • goat
  • mouse,..

Depends on choice of antibody (monoclonal requires mouse) and antibodies to be used in conjunction with antibody to be generated.

Type of antigen to use

  • full-length protein (natural selection of epitope, epitope location initially unknown, may not be availalbe, epitope may not be specific to protein)
  • peptide (cheaper, can typically be synthesised without problems, known epitope, possibly low antigenicity)

See also

External links

Protocol Online (methods forum)

Wikipedia

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