Antibody Production

Antibodies are proteins produced by the body in response to a foreign substance or antigen. An antigen can stimulate the production of a number of antibodies with specificity to different regions of the antigen. As antibody is produced by a single antibody forming cell, a clone and that an antigen can stimulate the development of several clones each secreting a distinct antibody, the pool of these antibodies in the blood is known as the polyclonal response. To isolate a single antibody producing cell, the technology known as lymphocyte hybridoma or monoclonal production is performed.

A STANDARD PROTOCOL INCLUDES:

• Synthesis and purification of peptide, including Mass specs and HPLC
  profiles

• Conjugation of peptide to a carrier protein as KLH, BSA, or the MAPS system

• 2 rabbits per peptide (6 months old, New Zealand, white female.)

Immunization protocol

Day 0: Preblood Initial immunization (200µg Antigen + adjuvant/rabbit)
Day 14: Boost with the same sample size as the initial injection.
Day 21: Boost the same as above.
Day 30: Test bleed and measure antibody's titer.
Day 60: Boost again if necessary.
Day 70: Test bleed and measure antibody's titer. We report the titer to the
customer and decide on the best option

*A minimum of 120 ml of crude serum will be delivered from the 2 rabbits

*Antibody titration with ELISA is performed with each bleeding

Sources Of Monoclonal Antibodies:
The starting material for monoclonal antibody production is antibody-producing cells known as lymphocytes which are isolated from the spleen of a rat or mouse that has been immunized with the antigen of interest. Since these lymphocytes have a limited life span, they are fused to malignant tumors called myelomas which can be propagated indefinitely in tissue culture. The fused cells known as lymphocyte hybridomas not only have the characteristics of the parent lymphocyte cell, that is, the ability to secrete specific antibody but also has acquired the myeloma's ability to
grow indefinitely.