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A human monoclonal anti-hepatitis B antibody preparation (TUVIRUMAB) was administered 6 times over a 2-week period in a dose-escalating scheme to chronic hepatitis B patients pre-treated with lamivudine. The capacity of the TUVIRUMAB antibody to "neutralize" hepatitis B surface antigen in the circulation was investigated by means of experimental enzyme-immunoassays. Monoclonal antibody conjugates enabled the detection of HBsAg, TUVIRUMAB, and HBsAg/TUVIRUMAB complexes. The results showed that (1) TUVIRUMAB was able partially to "neutralize" in vitro and in vivo, (2) HBsAg/TUVIRUMAB complexes can be traced by assays that capture the complex at either its HBsAg or its TUVIRUMAB component, (3) the final concentration of TUVIRUMAB at the end of therapy varied greatly but seemed to be related to HBsAg production at the start of therapy, (4) for at least 14 days after discontinuation of therapy, a minimal HBsAg level could be maintained in the presence of a declining TUVIRUMAB titer in patients with less than 3 microg/ml HBsAg before the start of therapy, (5) three months after therapy, all HBsAg levels had returned to pre-treatment levels and TUVIRUMAB had disappeared.

The feasibility to raise nonhuman primate antibodies against selected components of the human immune system was tested. The immunogens were whole cells (human T lymphocytes) or purified, recombinant human proteins (cytokines: TNF alpha or GM-CSF; soluble forms of cell surface antigens: sCD4 or sCD25). Significant immunizations, yielding functionally relevant antibodies, were readily achieved in rhesus monkeys, but, not surprisingly, may be less frequent in chimpanzees. The results suggest a general strategy for production of therapeutically useful MAB.

Animal cell technology is attracting considerable interest because of the capacity of animal cell cultures to synthesize or transform complex compounds such as virus vaccines, immunochemicals, hormones or enzymes. For the growth of surface-dependent cells, microcarrier technology is gaining importance. Here, we have attempted to immobilize surface-independent cells, normally grown in suspension, by entrapping them in polymer microbeads. Such entrapment should give increased stability to the normally fragile animal cells, allow for high cell densities to be achieved within the beads and make such preparations suitable for continuous operation. At the same time, the need for separation of the desired product from the cells is obviated. With the model systems studied, we showed that hybridoma, as well as other cell lines entrapped in agarose microbeads, remained viable. Both immunoglobulins and lymphokines were exported through the microbeads into the medium for 1-3 weeks, at levels corresponding well to those produced with free cells.