By Michael Heidelberger

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Theirs was a very fruitful collaboration of a pathologist and a biochemist. They postulated the theory in a vague way, and that is still, I think, the best way. If you try to draw too definite a picture of something you don't know much about, then you are pretty sure to be wrong. If you leave your description sufficiently vague, it may cover the actual facts. Breinl and Haurowitz postulated that antigen penetrated to the site of globulin formation, wherever that might be, and there so modified the course of globulin synthesis that there was a kind of complementarily shaped portion of the new protein that was formed, and that this new protein, because it had a complementary shape to the antigen or a portion of the antigen, could therefore combine with it when it met it in solution or in the blood stream at some future time.

Now, if one centrifuges off these pneumococci agglutinated by a large excess of antibody, and if one washes them well with salt solution until fresh pneumococci added to the supernatant solution are no longer agglutinated (that takes 3 or 4 thorough washings, centrifuging each time), one may then resuspend the washed cells and divide them into two parts. Let us say we have Type I pneumococci in an even suspension which does not change during some minutes and then very slowly begins to settle. One can show the specificity of the agglutination and the ability of the already agglutinated pneumococci to combine with antigen in multiple proportions by the experiment 56 which I shall now describe to you.

1 ) . Then one could calculate the amount of nitrogen to be precipitated from the ratio at the point of intersection of the perpendicular with the ratio line. Then, on this basis, we thought we would like to try a direct measurement of antibody nitrogen. W e began analyzing the pre- 38 cipitates for nitrogen and found the conditions under which we could wash out, within the limits of the micro-Kjeldahl method, all of the nonspecific nitrogen. When we had done this with the purified antibody solutions, we tried adding normal horse serum and found that we still could wash out all nonspecific protein.

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