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The expressions derived in the previous paper for the respective normal, F3, and shear forces, Fshear, acting along and perpendicular to the axis of a doublet of rigid spheres, were used to determine the hydrodynamic forces required to separate two red cell spheres of antigenic type B crosslinked by the corresponding antibody. Cells were sphered and swollen in isotonic buffered glycerol containing 8 X 10(-5) M sodium dodecyl sulfate, fixed in 0.085% glutaraldehyde, and suspended in aqueous glycerol (viscosity: 15-34 mPa s), containing 0.15 M NaCl and anti-B antibody from human hyperimmune antiserum at concentrations from 0.73 to 3.56 vol%. After incubating and mixing for 12 h, doublets were observed through a microscope flowing in a 178-micron tube by gravity feed between two reservoirs. Using a traveling microtube apparatus, the doublets were tracked in a constantly accelerating flow and the translational and rotational motions were recorded on videotape until breakup occurred. From a frame by frame replay of the tape, the radial position, velocity and orientation of the doublet were obtained and the normal and shear forces of separation at breakup computed. Both forces increased significantly with increasing antiserum concentration, the mean values of F3 increasing from 0.060 to 0.197 nN, and Fshear from 0.023 to 0.072 nN. There was no significant effect of glycerol viscosity on the forces of separation. It was not possible to determine whether the shear or normal force was responsible for doublet separation. Measurements of the mean dimensionless period of rotation, TG, of doublets in suspensions containing 0.73 and 2.40% antiserum undergoing steady flow were also made to test whether the spheres were rigidly linked or capable of some independent rotation. A fairly narrow distribution in TG about the value 15.64, predicted for rigidly-linked doublets, was obtained at both antiserum concentrations.

Carcinoembryonic antigen (CEA) was first detected by the immunization of rabbits with human colon cancer extract. The techniques of antiserum absorption and immunologic tolerance were employed in an attempt to render the resulting antisera tumor-specific. Using the procedures of precipitation and precipitin-inhibition in gel media, hemagglutination, passive cutaneous anaphylaxis and immunofluorescence, CEA was identified exclusively in all cancers of gastrointestinal origin and fetal digestive organs in the first two trimesters of gestation. The subsequent development of radioimmunoassays for CEA has raised the question of the presence of this material, in very low concentrations, in other normal and diseased (cancerous and noncancerous) tissue, but the problem of cross-reactivity within a family of closely related, but nonidentical, molecules remains to be resolved. CEA was initially purified by a sequence of steps involving extraction in perchloric acid, sieve chromatography, and preparative block electrophoresis. The molecule was characterized as a relatively heterogeneous acid glycoprotein with a molecular weight of approximately 200,000 and its carbohydrate (one-half to two-thirds of the molecule) and protein composition were determined. CEA was localized to the glycocalyx of the colon cancer cell and it was found that the CEA could be released from this site into the circulation of the cancer patient, where it could then be detected by means of radioimmunoassay. The clinical implications of this observation, as they have evolved over the past eight years, form the basis of the papers that constitute this supplement.

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