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US 12,577,296 B2Granted
Compositions and methods for growth factor modulation
AssigneeScholar Rock, Inc.
InventorsThomas Schurpf, Nagesh K. Mahanthappa, Michelle Marie Straub
Life of this patent
Estimating the term…
Abstract
Provided herein are proteins, antibodies, assays and methods useful for modulating growth factor levels and/or activities. In some embodiments, such growth factors are members of the TGF-β superfamily of proteins.
Claims11
A method for manufacturing a pharmaceutical composition that comprises an antibody, or an antigen-binding portion thereof, that inhibits transforming growth factor beta 1 (TGF-β1) signaling, the method comprising the steps of:
- (i) screening an antibody display library or hybridoma cell lines for an antibody, or an antigen-binding portion thereof, capable of binding an antigen comprising a transforming growth factor beta 1 prodomain complex (TGF-β1 GPC), that comprises:
- a) a latency associated peptide (LAP) dimer, wherein each polypeptide of the dimer has an amino acid sequence comprising SEQ ID NO:
- 38 or SEQ ID NO:
- 99;
- and
- b) a growth factor dimer, wherein each polypeptide of the dimer has an amino acid sequence comprising SEQ ID NO:
- 44;
- and
- selecting an antibody, or antigen-binding portion thereof, that binds to the TGF-β1 GPC;
- (ii) providing a growth factor activity assay to measure inhibition of TGF-β1 signaling comprising:
- a) contacting the TGF-β1 GPC with the antibody, or antigen-binding portion thereof, selected in step (i), a cell expressing αvβ6 integrin and/or αvβ8 integrin, and a responsive cell comprising a reporter gene comprising a TGF-β1 growth factor responsive promoter operably linked to a sequence encoding a detectable gene product;
- b) obtaining TGF-β1 signaling activity data by detecting expression of the detectable gene product, and
- c) selecting an antibody, or an antigen-binding portion thereof, that inhibits TGF-β1 signaling;
- (iii) preparing expression vectors comprising the coding sequences of the antibody, or the antigen-binding portions thereof, selected in step (ii);
- (iv) transfecting the vectors prepared in step (iii) into a cell in culture medium for expression of the antibody, or antigen-binding portion thereof;
- (v) purifying the antibody, or the antigen-binding portion thereof, expressed in step (iv) from the culture medium;
- and
- (vi) formulating the antibody, or the antigen-binding portion thereof, purified in step (v) into a pharmaceutical composition.
15 more elements
A method for manufacturing a pharmaceutical composition that comprises an antibody, or an antigen-binding portion thereof, that inhibits transforming growth factor beta 1 (TGF-β1) signaling, the method comprising the steps of:
- (i) screening an antibody display library or hybridoma cell lines for an antibody, or an antigen-binding portion thereof, capable of binding an antigen comprising a transforming growth factor beta 1 prodomain complex (TGF-β1 GPC), that comprises:
- a) a latency associated peptide (LAP) dimer, wherein each polypeptide of the dimer has an amino acid sequence comprising SEQ ID NO:
- 38 or SEQ ID NO:
- 99;
- and
- b) a growth factor dimer, wherein each polypeptide of the dimer has an amino acid sequence comprising SEQ ID NO:
- 44;
- and
- selecting an antibody, or antigen-binding portion thereof, that binds to the TGF-β1 GPC;
- (ii) providing a growth factor activity assay to measure inhibition of TGF-β1 signaling comprising:
- a) contacting the TGF-β1 GPC with the antibody, or antigen-binding portion thereof, selected in step (i), a cell expressing αvβ6 integrin and/or αvβ8 integrin, and a responsive cell comprising a reporter gene comprising a TGF-β1 growth factor responsive promoter operably linked to a sequence encoding a detectable gene product;
- b) obtaining TGF-β1 signaling activity data by detecting expression of the detectable gene product, and
- c) selecting an antibody, or an antigen-binding portion thereof, that inhibits TGF-β1 signaling;
- d) carrying out affinity maturation and/or optimization of the antibody, or antigen-binding portion thereof selected in step (ii) c);
- or
- humanizing the antibody, or the antigen-binding portion thereof selected in step (ii) c);
- or
- selecting an antibody, or antigen-binding portion thereof, from step (ii) c) that cross-reacts with human or mouse forms of the antigen;
- or
- performing an in vitro binding assay for the antibody, or antigen-binding portion thereof, selected in step (ii) c), and selecting an antibody, or antigen-binding portion thereof, that does not bind mature TGF-β1 growth factor or empty LAP dimer;
- (iii) preparing expression vectors comprising the coding sequences of the antibody, or the antigen-binding portions thereof, of step (ii) d);
- (iv) transfecting the vectors prepared in step (iii) into a cell in culture medium for expression of the antibody, or antigen-binding portion thereof;
- (v) purifying the antibody, or the antigen-binding portion thereof, expressed in step (iv) from the culture medium;
- and
- (vi) formulating the antibody, or the antigen-binding portion thereof, purified in step (v) into a pharmaceutical composition.
22 more elements
Dependencies are inferred from the claim text.