Production of Panels of Monoclonal Antibodies by the Hybridoma Method

  • Nguyen thi Man
  • Glenn E. Morris
Part of the Methods in Molecular Biology™ book series (MIMB, volume 66)


Since the first description by Kohler and Milstein (1), many variations on this method for the production of monoclonal antibodies (MAbs) have appeared (e.g., 2, 3, 4, and it may seem superfluous to add another. The variation we describe here, however, includes a number of refinements that enable rapid (6–10 wk) production from a single spleen of large numbers (20–30) of cloned, established hybridoma lines producing antibodies of high affinity. We have applied this method to recombinant fusion proteins containing fragments of the muscular dystrophy protein, dystrophin (5, 6, 7, 8) and dystrophin-related proteins (9,10); to hepatitis B surface antigen (11); and to the enzyme, creatine kinase (12). We have used the MAbs thus produced for immunodiagnosis, epitope mapping, and studies of protein structure and function (5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15). Epitopes shared with other proteins are common (e.g., dystrophin and α-actinin [16]), so availability of several MAbs against different epitopes on a protein can be important in ensuring the desired specificity in immunolocalization and Western blotting studies (9).


Creatine Kinase Myeloma Cell Microwell Plate Western Blotting Study Resuspend Pellet 
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Copyright information

© Humana Press Inc. 1996

Authors and Affiliations

  • Nguyen thi Man
    • 1
  • Glenn E. Morris
    • 2
  1. 1.Research DivisionNorth East Wales InstituteWrexhamUK
  2. 2.MRIC Biotechnology GroupNorth East Wales InstituteWrexhamUK

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