Appeal No. 1998-1987
Application No. 07/915,783
antibodies. See page 212, right-hand column. Meanwhile, Plasmodium knowlesi
antigen was prepared by extracting parasite-infected red blood cells by
“suspend[ing] in 1 ml PBS [phosphate-buffered saline] containing 1% Triton X-
100 . . . [and] incubat[ing] on ice for 1 hr with intermittent vigorous vortexing.”
Page 213, paragraph bridging the columns.4 The resulting soluble extract was
“mixed with ascites fluid containing the monoclonal antibody.”5 Id., right-hand
column. After the monoclonal antibody had been allowed to bind the plasmodial
antigens in the extract, “goat anti-mouse Ig-conjugated Sepharose 4B or protein
A-Sepharose 4B” was added. After the immune complexes (i.e., plasmodial
antigen plus monoclonal antibody) had been allowed to bind, the Sepharose gels
were washed to eliminate unbound extract and “[i]mmune complexes were eluted
from the Sepharose.” Id.
Thus, both references disclose compositions comprising Sepharose beads
with protein A attached and an antibody/plasmodial antigen immune complex
bound to the protein A. These compositions are water-insoluble, since both
Kilejian and Epstein recover the Sepharose-containing composition by
centrifugation. See Kilejian at page 3696 (“[Sepharose] beads were pelleted and
washed once with the buffer. . . . Unbound extract was removed by three washes
with 0.5 M LiCl/10 mM Tris-HCl, pH 8, and one wash with 1% Nonidet P-40
buffer.”); Epstein at page 213 (“[Sepharose] gels with bound immune complexes
were washed twice with 5 ml [NETT] buffer. . . containing 10% FBS, followed by
3 Nonidet P-40 is a non-ionic detergent. See, e.g., claim 18.
4 Triton X-100 is a non-ionic detergent. See, e.g., claim 18.
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