Claims
- 1. An isolated protein having an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 1-69 and 139-181.
- 2. The isolated protein of claim 1, wherein said protein is an antigen-binding protein.
- 3. The isolated protein of claim 2, wherein said antigen is human Rh(D) protein.
- 4. The isolated protein of claim 3, wherein said protein has an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-69 and 139-181.
- 5. The isolated protein of claim 3, wherein said antigen-binding protein is an antibody.
- 6. The isolated protein of claim 5, wherein said antibody comprises a heavy chain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-28 and 139-153.
- 7. The isolated protein of claim 5, wherein said antibody comprises a light chain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 29-69 and 154-181.
- 8. The isolated protein of claim 5, wherein said antibody comprises a heavy chain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-28 and 139-153 and a light chain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 29-69 and 154-181.
- 9. The isolated protein of claim 3, wherein said binding protein is an antibody fusion protein.
- 10. The isolated protein of claim 1, wherein said protein is substantially purified.
- 11. An isolated DNA encoding a protein having an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 1-69 and 139-181.
- 12. The isolated DNA of claim 10, having a nucleotide sequence selected from the group consisting of SEQ ID NOs: 70-138 and 182-224.
- 13. The isolated DNA of claim 12, being substantially purified.
- 14. An isolated DNA encoding a protein obtained by
generating a synthetic DNA library in a virus vector expressing said protein; adding a magnetic label to cells expressing said antigen-bearing moiety; incubating virus expressing said protein with said magnetically labeled cells in the presence of an excess of non-labeled cells which do not express said antigen-bearing moiety to form a mixture, wherein said virus binds to said magnetically labeled cells; isolating virus bound cells from said mixture and obtaining DNA encoding said protein therefrom.
- 15. The isolated DNA of claim 14, having a nucleotide sequence selected from the group consisting of SEQ ID NOs: 70-138 and 182-224.
- 16. A substantially pure protein obtained by
generating a synthetic DNA library in a virus vector expressing said protein; adding a magnetic label to cells expressing said antigen-bearing moiety; incubating virus expressing said protein with said magnetically labeled cells in the presence of an excess of non-labeled cells which do not express said antigen-bearing moiety to form a mixture, wherein said virus binds to said magnetically labeled cells; isolating virus bound cells from said mixture and isolating said protein therefrom.
- 17. The substantially pure protein of claim 16, having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-69 and 139-181.
- 18. A substantially pure preparation of a protein obtained by expressing said protein from DNA encoding said protein, wherein said DNA is obtained by
generating a synthetic DNA library in a virus vector expressing said protein; adding a magnetic label to cells expressing said antigen-bearing moiety; incubating virus expressing said protein with said magnetically labeled cells in the presence of an excess of non-labeled cells which do not express said antigen-bearing moiety to form a mixture, wherein said virus binds to said magnetically labeled cells; isolating virus bound cells from said mixture and obtaining DNA encoding said protein therefrom.
- 19. The substantially pure protein of claim 18, having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-69 and 139-181.
- 20. A method of isolating a DNA encoding a multi-subunit protein which binds to an antigen-bearing moiety, said method comprising
generating a phage display library comprising a plurality of virus vectors, wherein a first of said virus vectors comprises a first heterologous DNA encoding a subunit of said protein and expresses said subunit on the surface thereof, and wherein a second of said virus vectors comprises a second heterologous DNA encoding a different subunit of said protein and expresses said different subunit on the surface thereof; adding a magnetic label to cells bearing said antigen-bearing moiety on their surface; incubating said phage display library with said magnetically labeled cells in the presence of an excess of non-labeled cells which do not express said antigen-bearing moiety to form a mixture, whereby said first and second virus vectors bind to said magnetically labeled cells; isolating magnetically labeled cells from said mixture, whereby said first and second virus vectors are isolated from said mixture; obtaining said first heterologous DNA from said first virus vector; ligating at least the portion of said first heterologous DNA encoding said subunit and at least the portion of said second heterologous DNA encoding said different subunit to form a hybrid heterologous DNA; generating a hybrid virus vector comprising said hybrid heterologous DNA and expressing said subunit and said different subunit of said protein on the surface thereof; adding a magnetic label to cells bearing said antigen-bearing moiety on their surface; incubating said hybrid virus vector with said magnetically labeled cells in the presence of an excess of non-labeled cells which do not express said antigen-bearing moiety to form a mixture, whereby said hybrid virus vector binds to said magnetically labeled cells; isolating magnetically labeled cells from said mixture, whereby said hybrid virus vector is isolated from said mixture; and obtaining DNA encoding said protein from said isolated virus vector, whereby said DNA is isolated.
- 21. A method of isolating a multi-subunit protein which binds to an antigen-bearing moiety, said method comprising
generating a phage display library comprising a plurality of virus vectors, wherein a first of said virus vectors comprises a first heterologous DNA encoding a subunit of said protein and expresses said subunit on the surface thereof, and wherein a second of said virus vectors comprises a second heterologous DNA encoding a different subunit of said protein and expresses said different subunit on the surface thereof; adding a magnetic label to cells bearing said antigen-bearing moiety on their surface; incubating said phage display library with said magnetically labeled cells in the presence of an excess of non-labeled cells which do not express said antigen-bearing moiety to form a mixture, whereby said first and second virus vectors bind to said magnetically labeled cells; isolating magnetically labeled cells from said mixture, whereby said first and second virus vectors are isolated from said mixture; obtaining said first heterologous DNA from said first virus vector; ligating at least the portion of said first heterologous DNA encoding said subunit and at least the portion of said second heterologous DNA encoding said different subunit to form a hybrid heterologous DNA; generating a hybrid virus vector comprising said hybrid heterologous DNA and expressing said subunit and said different subunit of said protein on the surface thereof; adding a magnetic label to cells bearing said antigen-bearing moiety on their surface; incubating said hybrid virus vector with said magnetically labeled cells in the presence of an excess of non-labeled cells which do not express said antigen-bearing moiety to form a mixture, whereby said hybrid virus vector binds to said magnetically labeled cells; isolating magnetically labeled cells from said mixture, whereby said hybrid virus vector is isolated from said mixture; and isolating said protein from said isolated virus vector, whereby said protein is isolated.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is entitled to priority pursuant to 35 U.S.C. §119(e) to U.S. provisional patent application No. 60/081,380, which was filed on Apr. 10, 1998, and is a continuation-in-part of U.S. application Ser. No. 08/884,045, filed Jun. 27, 1997, which application is entitled to priority under 35 U.S.C. §119(e) to U.S. Provisional Application No. 60/028,550, filed on Oct. 11, 1996.
GOVERNMENT SUPPORT
[0002] This invention was supported in part by a grant from the U.S. Government (NIH Grant No. P50-HL54516) and the U.S. Government may therefore have certain rights in the invention.
Provisional Applications (2)
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Number |
Date |
Country |
|
60081380 |
Apr 1998 |
US |
|
60028550 |
Oct 1996 |
US |
Divisions (1)
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Number |
Date |
Country |
Parent |
09240274 |
Jan 1999 |
US |
Child |
09848798 |
May 2001 |
US |
Continuation in Parts (1)
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Number |
Date |
Country |
Parent |
08884045 |
Jun 1997 |
US |
Child |
09240274 |
Jan 1999 |
US |