Sri International

Nippon Mining Co., Ltd.

Paul H Johnson

Jerome B Lazar

Nahid S Waleh

Christopher J Green

Richard C Winant

David Gray Streeter

Indira Sohel

Ping Sze

The present invention provides for the purification from native sources or the cloning and expression of a variant form of platelet factor 4 and also provides recombinant DNA vectors and methods for the expression and recovery of the platelet factor 4 variant. Also provided are compositions and methods for modulating immune responses in mammals comprising immunomodulating effective amounts of platelet factor 4 variant.

Cloning and expression of a variant gene of platelet factor 4 and

The present invention provides a process for the recovery of heterologous proteins from CTAP-III fusion proteins comprising expressing a fusion protein having a first amino acid sequence, a second amino acid sequence, and a selectable site which may be cleaved to provide first and second polypeptide fragments, respectively, wherein the first amino acid fragment is homologous to CTAP-III, and the first and second fragments have different pI values; cleaving the fusion protein to provide the first and second fragments; and separating the first and second fragments by ion exchange chromatography.

Purification of proteins employing CTAP-III fusions

Human laminin B.sub.1 peptides have been produced. The invention provides novel peptides which contain substantial amino acid sequence similarity to amino acids 897 to 936 in the human sequence. These peptides have been found to have enhanced antimetastatic activity.

Antimetastatic peptides with laminin activity

A method for obtaining heterologous peptides from fusion proteins wherein at least one of the fusion components is connective tissue-activating peptide-III is provided. Hirudin, a laminin B.sub.1 peptide and platelet factor 4 are polypeptides expressed using this method. DNA sequences encoding the fusion protein, vectors containing these sequences and transformed prokaryotic hosts useful in practicing the method of the present invention are also provided.

Method and vectors for stabilizing hirudin and human laminin B.sub.1

Compounds and compositions comprising amino acid substituted and chemically modified analogs of hirudin are provided, together with methods for their production and use as antithrombotic and anticoagulant agents.

Analogs of hirudin

The present invention provides a method to improve the expression of a poorly expressed recombinant protein, such as platelet factor 4, by constructing a gene fusion such that unfavorable secondary structure formed between the region of the messenger RNA specifying the recombinant protein and the region of the messenger RNA containing the ribosome binding site is disrupted. The gene fusion is constructed by inserting a DNA segment of sufficient length to disrupt unfavorable long-range folding interactions which render the ribosome binding site inaccesible to ribosomes thereby interfering with the translation of mRNA. The invention further provides for the expression of recombinant platelet factor 4 and provides recombinant DNA vectors and methods for the bacterial production and recovery of said platelet factor 4.

Method to improve mRNA translation and use thereof for production of

A synthetic structural gene encoding CTAP-III or CTAP-III-Leu21 including adaptors for the carboxy and amino terminal ends of the gene which contain start and stop codons and convenient restriction sites for use in cloning the gene is described. The gene was designed for efficient expression in bacteria and to include two unique restriction sites for BamHI and XbaI. Plasmid expression vectors that are derivatives of pBR322 that contain a ColE1 insert which includes the expression control sequence and structural gene for colicin are also described. Constructs for expressing CTAP-III, CTAP-III-Leu21, CTAP-III or CTAP-III-Leu21 having a nonpolar pentapeptide fused to its amino terminus, and fusion proteins of such CTAP-III proteins and a colicin fragment are prepared by inserting the synthetic structural gene into the vectors at positions in phase with the colicin expression control sequence and under the control thereof.

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Menlo Park, CA, United States of America

Paul H Johnson