A General Method of Making Antibodies (Antitoxins, Immune Sera, Precipitins, Agglutinins) Outside of the Animal Body. Linus Pauling. May 19, 1941.

In a paper entitled "Theory of the Structure and Process of Formation of Antibodies, " Linus Pauling, Jour. Amer. Chem. Soc. 62, 2643 (1940), there was suggested a general method of producing antibodies differing from the methods used at present, which consist in the production of serum by a suitably treated animal. The pertinent paragraph is the following (page 2656):

"An interesting possible method of producing antibodies from serum or globulin solution outside of the animal is suggested by the theory. The globulin would be treated with a denaturing agent or condition sufficiently strong to cause the chain ends to uncoil; after which this agent or condition would be removed slowly while antigen or hapten is present in the solution in considerable concentration. The chain ends would then coil up to assume the configurations stable under these conditions which would be configurations complementary to those of the antigen or hapten."

The material to be converted into the antibody solution could be normal serum from some animal, some fraction of this (a globulin or albumen), or another suitable protein solution. The denaturing agent or condition could be increased temperature, change in pH (treatment with base or acid), or some substance such as urea or salicylates, or some combination of these. As antigen there might be used a bacterium or bacillus, a toxin, a synthetic hapten, or other suitable substance.

Experiments have been carried out so far with three different fractions of beef globulin as raw material and with a dye as antigen. Both increase in temperature to about 50° C. and increase in pH to about 10.7 were used separately as denaturing conditions or agents. With both conditions slow renaturation in the presence of the antigen resulted in the production of substances having the characteristic properties of antibodies specific to the antigen. Various check experiments with conditions varied have also been carried out.

I think that there it the possibility that this general method may be of practical value in the manufacture of certain antitoxins and other other sera.