SHBG AND IBP4 ANTIBODIES AND METHODS OF USING SAME

Abstract

Anti-SHBG and anti-IBP4 antibodies or antigen-binding fragments thereof are described. Nucleic acids encoding the antibodies, compositions comprising the antibodies, methods of producing the antibodies, and methods of using the antibodies are also described. The present disclosure relates to anti-sex hormone-binding globulin (SHBG) antibodies and anti-insulin-like growth factor binding protein 4 (IBP4), nucleic acids and expression vectors encoding the antibodies, recombinant cells containing the vectors, and compositions comprising the antibodies. The present disclosure further relates to methods of making the antibodies and methods of using the antibodies to diagnose or treat conditions associated with SHBG and / or IBP4.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 63 / 308,453, filed February 9, 2022, the entire contents of which are incorporated herein by reference.

[0002] Sequence Listing This application contains a Sequence Listing that was submitted via the Patent Center. The Sequence Listing, entitled 203123-025002PCT.xml, created on February 9, 2023, and having a size of 320,751 bytes, is hereby incorporated by reference in its entirety.

[0003] FIELD OF THE INVENTION The present invention relates to anti-sex hormone-binding globulin (SHBG) antibodies and anti-insulin-like growth factor-binding protein 4 (IBP4), nucleic acids and expression vectors encoding the antibodies, recombinant cells containing the vectors, and compositions comprising the antibodies. Methods of making the antibodies and using the antibodies to diagnose preterm labor or pregnancy-related complications are also provided. [Background technology]

[0004] Background of the Invention background According to the World Health Organization, an estimated 15 million babies are born prematurely (before 37 weeks of gestation) each year. Premature birth rates are increasing in almost every country with reliable data. World Health Organization; March of Dimes; The Partnership for Maternal, Newborn & Child Health; Save the See Children, Born Too Soon: The Global Action Report on Preterm Birth, ISBN 9789241503433 (2012). An estimated 1 million infants die each year from complications of preterm birth. Globally, premature birth is the number one cause of death in newborns (infants in the first four weeks of life) and the second leading cause of death in children under five, after pneumonia. Many survivors face a lifetime of disabilities, including learning disabilities and vision and hearing problems.

[0005] To prevent preterm birth in women less than 24 weeks pregnant who have an open cervix on ultrasound, a surgical procedure known as cervical cerclage can be used to close the cervix with strong sutures. Women less than 34 weeks pregnant and in active preterm labor may require hospitalization and the administration of medications to temporarily stop preterm labor and / or promote fetal lung development. If a pregnant woman is determined to be at risk for preterm labor, healthcare providers can implement a variety of clinical strategies, including preventative oral medications, such as 17α-hydroxyprogesterone caproate (Makena) injections and / or vaginal progesterone gel, cervical pessaries, restrictions on sexual and / or other physical activities, and changes in treatment for chronic conditions that increase the risk of preterm labor, such as diabetes and hypertension.

[0006] There is a significant need to identify women at risk for preterm birth and provide them with appropriate prenatal care. Women identified as high-risk can be planned for more intensive prenatal surveillance and preventive interventions. Current strategies for risk assessment are based on obstetric and medical histories and clinical examinations, but these strategies only identify a small percentage of women at risk for preterm birth. Currently, a prior history of spontaneous preterm birth (sPTB) is the single strongest predictor of subsequent preterm birth (PTB). A previous sPTB increases the likelihood of a second PTB by 30–50%. Other maternal risk factors include black race, low maternal body mass index, and short cervical length. Studies of amniotic fluid, cervicovaginal fluid, and serum biomarkers to predict sPTB suggest that abnormalities in multiple molecular pathways exist in women who ultimately deliver preterm. Reliable early identification of risk for preterm birth would allow for appropriate monitoring and clinical management planning to prevent preterm birth. Such monitoring and management may include more frequent prenatal visits, serial cervical length measurements, increased education about the signs and symptoms of early preterm labor, lifestyle interventions for modifiable risk behaviors, smoking cessation, cervical pessaries, and progesterone treatment. Finally, reliable prenatal identification of risk for preterm birth is also crucial for the cost-effective allocation of monitoring resources. Despite intensive research into identifying at-risk women, PTB prediction algorithms based solely on clinical and demographic factors or using serum or vaginal biomarkers have not yet led to clinically useful tests.To enable clinical intervention, more accurate methods are needed to identify at-risk women during their first and early pregnancy.The present disclosure addresses this need by providing SHBG and IBP4 binding agents for the detection and quantification of biomarkers that indicate susceptibility to preterm birth. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] March of Dimes;The Partnership for Maternal, Newborn & Child Health; Save the Children, Born too soon: the global action report on preterm birth, ISBN9789241503433 (2012) Summary of the Invention [Means for solving the problem]

[0008] A brief summary of the invention The present disclosure relates to anti-sex hormone-binding globulin (SHBG) antibodies and anti-insulin-like growth factor-binding protein 4 (IBP4), nucleic acids and expression vectors encoding the antibodies, recombinant cells containing the vectors, and compositions comprising the antibodies. The disclosure further relates to methods of making the antibodies and methods of using the antibodies to diagnose or treat conditions associated with SHBG and / or IBP4.

[0009] In one aspect, the disclosure provides an anti-SHBG antibody or fragment thereof, comprising: (i) a heavy chain complementarity determining region 1 (HCDR1) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 11, 14, 17, 20, 52, and 115; and a heavy chain complementarity determining region 2 (HCDR2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117; and an amino acid sequence selected from the group consisting of SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54. or (ii) a heavy chain comprising a heavy chain complementarity determining region 3 (HCDR3) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23, 26, 29, and 32; or (ii) a light chain complementarity determining region 1 (LCDR1) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23, 26, 29, and 32; and a light chain complementarity determining region 2 (LCDR2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 24, 27, 30, and 33; and a light chain complementarity determining region 3 (LCDR3) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 25, 31, and 34. In some embodiments, the isolated anti-SHBG antibody or fragment thereof comprises a heavy chain. In some embodiments, the isolated anti-SHBG antibody or fragment thereof comprises a light chain. In some embodiments, the isolated anti-SHBG antibody or fragment thereof comprises a heavy chain and a light chain.

[0010] In some embodiments, the disclosure provides an anti-SHBG antibody or fragment thereof, wherein HCDR1 comprises the amino acid sequence of SEQ ID NO: 11, HCDR2 comprises the amino acid sequence of SEQ ID NO: 12, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 13; and the light chain comprises LCDR1 comprising the amino acid sequence of SEQ ID NO: 23, LCDR2 comprises the amino acid sequence of SEQ ID NO: 24, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO: 11, HCDR2 comprises the amino acid sequence of SEQ ID NO: 44, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 45; LCDR1 comprises the amino acid sequence of SEQ ID NO: 23, LCDR2 comprises the amino acid sequence of SEQ ID NO: 24, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO: 14, HCDR2 comprises the amino acid sequence of SEQ ID NO: 15, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 16; LCDR1 comprises the amino acid sequence of SEQ ID NO: 26, LCDR2 comprises the amino acid sequence of SEQ ID NO: 27, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO: 14, HCDR2 comprises the amino acid sequence of SEQ ID NO: 47, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 48; LCDR1 comprises the amino acid sequence of SEQ ID NO: 26, LCDR2 comprises the amino acid sequence of SEQ ID NO: 27, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO: 17, HCDR2 comprises the amino acid sequence of SEQ ID NO: 18, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 19; LCDR1 comprises the amino acid sequence of SEQ ID NO: 29, LCDR2 comprises the amino acid sequence of SEQ ID NO: 30, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 31. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO: 17, HCDR2 comprises the amino acid sequence of SEQ ID NO: 50, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 51; LCDR1 comprises the amino acid sequence of SEQ ID NO: 29, LCDR2 comprises the amino acid sequence of SEQ ID NO: 30, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 31.In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:20, HCDR2 comprises the amino acid sequence of SEQ ID NO:21, and HCDR3 comprises the amino acid sequence of SEQ ID NO:22; LCDR1 comprises the amino acid sequence of SEQ ID NO:32, LCDR2 comprises the amino acid sequence of SEQ ID NO:33, and LCDR3 comprises the amino acid sequence of SEQ ID NO:34. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:52, HCDR2 comprises the amino acid sequence of SEQ ID NO:53, and HCDR3 comprises the amino acid sequence of SEQ ID NO:54; LCDR1 comprises the amino acid sequence of SEQ ID NO:32, LCDR2 comprises the amino acid sequence of SEQ ID NO:33, and LCDR3 comprises the amino acid sequence of SEQ ID NO:34. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO:115, HCDR2 comprises the amino acid sequence of SEQ ID NO:116, and HCDR3 comprises the amino acid sequence of SEQ ID NO:19; LCDR1 comprises the amino acid sequence of SEQ ID NO:29, LCDR2 comprises the amino acid sequence of SEQ ID NO:30, and LCDR3 comprises the amino acid sequence of SEQ ID NO:31. In some embodiments, HCDR1 comprises the amino acid sequence of SEQ ID NO: 115, HCDR2 comprises the amino acid sequence of SEQ ID NO: 116, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 51; LCDR1 comprises the amino acid sequence of SEQ ID NO: 29, LCDR2 comprises the amino acid sequence of SEQ ID NO: 30, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 31.

[0011] In another aspect, the disclosure provides an isolated anti-SHBG antibody or fragment thereof, comprising: (i) a heavy chain comprising an HCDR1 comprising the amino acid sequence GYTFTGYW (SEQ ID NO: 11), an HCDR2 comprising the amino acid sequence IDPSX1X2YT (SEQ ID NO: 112), where X1 is independently any naturally occurring amino acid residue and X2 is independently any naturally occurring amino acid residue, and an HCDR3 comprising the amino acid sequence TRX3PLVTADFX3Y (SEQ ID NO: 113), where X3 is independently any naturally occurring amino acid residue; and (ii) a light chain comprising an LCDR1 comprising the amino acid sequence QNVDTN (SEQ ID NO: 23), an LCDR2 comprising the amino acid sequence SAS (SEQ ID NO: 24), and an LCDR3 comprising the amino acid sequence QQYDSYPY (SEQ ID NO: 25). In some embodiments, X1 comprises threonine or asparagine, X2 comprises glycine or threonine, and X3 comprises glutamic acid or asparagine. In some embodiments, X1 comprises threonine, X2 comprises glycine, and X3 comprises glutamic acid.

[0012] In another aspect, the disclosure provides an isolated anti-SHBG antibody or fragment thereof, comprising: (i) a heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:4, or a light chain comprising an amino acid sequence at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:6; or (ii) a heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:36, or a light chain comprising an amino acid sequence at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:38. In some embodiments, the antibody or fragment thereof comprises the heavy chain of SEQ ID NO:4. In some embodiments, the antibody or fragment thereof of claim 18 comprises a light chain of SEQ ID NO: 6. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 4 and a light chain of SEQ ID NO: 6. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 36. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 38. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 36 and a light chain of SEQ ID NO: 38. In some embodiments, the heavy and light chains form an antigen-binding domain for binding to an epitope of SHBG. In some embodiments, the epitope of SHBG comprises the amino acid sequence of SEQ ID NO: 110.

[0013] In some embodiments, the present disclosure provides an isolated anti-SHBG antibody or fragment thereof, which has a higher affinity for a biologically active form of SHBG than for a biologically inactive form of SHBG. In some embodiments, the antibody or fragment thereof comprises a higher affinity for glycosylated SHBG than for non-glycosylated SHBG. In some embodiments, the glycosylated SHBG comprises an epitope comprising a glycosylated amino acid, the glycosylated amino acid comprising threonine at position 36. In some embodiments, the glycosylated amino acid comprises an O-linked glycan.

[0014] In another aspect, the disclosure provides an isolated anti-IBP4 antibody or fragment thereof, comprising: (i) a heavy chain complementarity determining region 1 (HCDR1) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 284, and 314; and SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 288, 291, 284, and 314. heavy chain complementarity determining region 2 (HCDR2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322; or (ii) a heavy chain comprising a heavy chain complementarity determining region 3 (HCDR3) comprising the amino acid sequence of SEQ ID NOs: 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330; and a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300 , 302, 326, 328, and 331; and a light chain comprising a light chain complementarity determining region 2 (LCDR2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332. In some embodiments, the antibody or fragment thereof comprises a heavy chain. In some embodiments, the antibody or fragment thereof comprises a light chain. In some embodiments, the antibody or fragment thereof comprises a heavy chain and a light chain.

[0015] In some embodiments, the present disclosure provides an isolated anti-IBP4 antibody or fragment thereof, wherein (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 75, HCDR2 comprises the amino acid sequence of SEQ ID NO: 76, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 77; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 87, LCDR2 comprises the amino acid sequence of SEQ ID NO: 24, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 99. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 78, HCDR2 comprises the amino acid sequence of SEQ ID NO: 79, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 80; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 90, LCDR2 comprises the amino acid sequence of SEQ ID NO: 91, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 89. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 78, HCDR2 comprises the amino acid sequence of SEQ ID NO: 79, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 80; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 90, LCDR2 comprises the amino acid sequence of SEQ ID NO: 91, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 99. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 81, HCDR2 comprises the amino acid sequence of SEQ ID NO: 82, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 83; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 93, LCDR2 comprises the amino acid sequence of SEQ ID NO: 94, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 95. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 81, HCDR2 comprises the amino acid sequence of SEQ ID NO: 82, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 83; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 93, LCDR2 comprises the amino acid sequence of SEQ ID NO: 94, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 100.In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 84, HCDR2 comprises the amino acid sequence of SEQ ID NO: 85, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 86; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 96, LCDR2 comprises the amino acid sequence of SEQ ID NO: 97, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 98. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 84, HCDR2 comprises the amino acid sequence of SEQ ID NO: 85, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 86; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 96, LCDR2 comprises the amino acid sequence of SEQ ID NO: 97, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 101. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 118, HCDR2 comprises the amino acid sequence of SEQ ID NO: 119, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 83; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 93, LCDR2 comprises the amino acid sequence of SEQ ID NO: 94, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 89. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 118, HCDR2 comprises the amino acid sequence of SEQ ID NO: 119, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 83; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 93, LCDR2 comprises the amino acid sequence of SEQ ID NO: 94, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 99.

[0016] In some embodiments, the present disclosure provides an anti-IBP4 antibody or fragment thereof, wherein the heavy and light chains form an antigen-binding domain for binding to an epitope of IBP4. In some embodiments, the epitope of IBP4 comprises the amino acid sequence of SEQ ID NO: 111.

[0017] In some embodiments, the present disclosure provides an isolated anti-IBP4 antibody or fragment thereof, wherein (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 75, HCDR2 comprises the amino acid sequence of SEQ ID NO: 216, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 217; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 226, LCDR2 comprises the amino acid sequence of SEQ ID NO: 24, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 227. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 78, HCDR2 comprises the amino acid sequence of SEQ ID NO: 218, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 219; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 228, LCDR2 comprises the amino acid sequence of SEQ ID NO: 229, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 227. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 81, HCDR2 comprises the amino acid sequence of SEQ ID NO: 220, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 221; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 230, LCDR2 comprises the amino acid sequence of SEQ ID NO: 231, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 232. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 84, HCDR2 comprises the amino acid sequence of SEQ ID NO: 223, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 223; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 233, LCDR2 comprises the amino acid sequence of SEQ ID NO: 234, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 235. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 118, HCDR2 comprises the amino acid sequence of SEQ ID NO: 224, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 225; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 230, LCDR2 comprises the amino acid sequence of SEQ ID NO: 231, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 227.In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 244, HCDR2 comprises the amino acid sequence of SEQ ID NO: 245, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 246; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 87, LCDR2 comprises the amino acid sequence of SEQ ID NO: 24, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 259. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 247, HCDR2 comprises the amino acid sequence of SEQ ID NO: 248, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 249; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 90, LCDR2 comprises the amino acid sequence of SEQ ID NO: 260, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 259. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 250, HCDR2 comprises the amino acid sequence of SEQ ID NO: 251, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 252; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 93, LCDR2 comprises the amino acid sequence of SEQ ID NO: 261, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 262. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 253, HCDR2 comprises the amino acid sequence of SEQ ID NO: 254, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 255; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 96, LCDR2 comprises the amino acid sequence of SEQ ID NO: 263, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 264. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 256, HCDR2 comprises the amino acid sequence of SEQ ID NO: 257, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 258; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 93, LCDR2 comprises the amino acid sequence of SEQ ID NO: 261, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 259. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 244, HCDR2 comprises the amino acid sequence of SEQ ID NO: 245, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 246; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 23, LCDR2 comprises the amino acid sequence of SEQ ID NO: 24, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 259.In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 247, HCDR2 comprises the amino acid sequence of SEQ ID NO: 248, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 249; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 272, LCDR2 comprises the amino acid sequence of SEQ ID NO: 273, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 259. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 250, HCDR2 comprises the amino acid sequence of SEQ ID NO: 251, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 252; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 274, LCDR2 comprises the amino acid sequence of SEQ ID NO: 94, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 262. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 253, HCDR2 comprises the amino acid sequence of SEQ ID NO: 254, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 255; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 275, LCDR2 comprises the amino acid sequence of SEQ ID NO: 97, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 264. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 256, HCDR2 comprises the amino acid sequence of SEQ ID NO: 257, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 258; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 274, LCDR2 comprises the amino acid sequence of SEQ ID NO: 94, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 259. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 244, HCDR2 comprises the amino acid sequence of SEQ ID NO: 284, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 285; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 87, LCDR2 comprises the amino acid sequence of SEQ ID NO: 297, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 298. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 247, HCDR2 comprises the amino acid sequence of SEQ ID NO: 286, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 287; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 90, LCDR2 comprises the amino acid sequence of SEQ ID NO: 299, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 298.In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 288, HCDR2 comprises the amino acid sequence of SEQ ID NO: 289, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 290; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 93, LCDR2 comprises the amino acid sequence of SEQ ID NO: 300, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 301. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 291, HCDR2 comprises the amino acid sequence of SEQ ID NO: 292, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 293; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 96, LCDR2 comprises the amino acid sequence of SEQ ID NO: 302, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 303. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 294, HCDR2 comprises the amino acid sequence of SEQ ID NO: 295, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 296; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 93, LCDR2 comprises the amino acid sequence of SEQ ID NO: 300, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 298. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 244, HCDR2 comprises the amino acid sequence of SEQ ID NO: 312, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 313; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 323, LCDR2 comprises the amino acid sequence of SEQ ID NO: 24, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 324. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 314, HCDR2 comprises the amino acid sequence of SEQ ID NO: 315, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 316; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 325, LCDR2 comprises the amino acid sequence of SEQ ID NO: 326, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 324. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 288, HCDR2 comprises the amino acid sequence of SEQ ID NO: 317, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 318; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 327, LCDR2 comprises the amino acid sequence of SEQ ID NO: 328, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 329.In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 291, HCDR2 comprises the amino acid sequence of SEQ ID NO: 319, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 320; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 330, LCDR2 comprises the amino acid sequence of SEQ ID NO: 331, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 332. In some embodiments, (i) HCDR1 comprises the amino acid sequence of SEQ ID NO: 294, HCDR2 comprises the amino acid sequence of SEQ ID NO: 321, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 322; (ii) LCDR1 comprises the amino acid sequence of SEQ ID NO: 327, LCDR2 comprises the amino acid sequence of SEQ ID NO: 328, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 324.

[0018] In another aspect, the present disclosure provides an isolated anti-IBP4 antibody or fragment thereof, comprising: (i) an HCDR1 comprising the amino acid sequence GYTFTDYY (SEQ ID NO: 75), an HCDR2 comprising the amino acid sequence INPNNGGS (SEQ ID NO: 76), and an HCDR3 comprising the amino acid sequence ARWDGGFDY (SEQ ID NO: 77); and (ii) an LCDR1 comprising the amino acid sequence QNVGTN (SEQ ID NO: 87), an LCDR2 comprising the amino acid sequence SAS (SEQ ID NO: 24), and an LCDR3 comprising the amino acid sequence QQYNX1YPLT (SEQ ID NO: 114), where X1 is any naturally occurring amino acid residue. In a further embodiment, X1 comprises isoleucine or serine.

[0019] In another aspect, the disclosure provides an isolated anti-IBP4 antibody or fragment thereof, comprising: (i) an HCDR1 comprising the amino acid sequence GYTFTYX1 (SEQ ID NO: 354), an HCDR2 comprising the amino acid sequence IX2X3X4X5GGX6 (SEQ ID NO: 355), and an HCDR3 comprising the amino acid sequence X7RX8X9X 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 DX 18(ii) a heavy chain comprising an HCDR3 comprising the amino acid sequence QNVX 19 X 20 X 21 LCDR1 comprising (SEQ ID NO: 357), amino acid sequence SX 22 LCDR2 containing S (SEQ ID NO: 358), and the amino acid sequence QQYX 23 X 24 YPLX 25 (SEQ ID NO: 359), wherein X1 to X5, X7, X 12 ~X 14 , X 18 ~X 20 , X 22 , and X 24 ~X 25 are independently any naturally occurring amino acid residue, X6 is independently any naturally occurring amino acid residue except isoleucine, X8 is independently a deletion or any naturally occurring amino acid residue except asparagine, X9 is independently any naturally occurring amino acid residue except tyrosine, and X 10 are independently a deletion or any naturally occurring amino acid residue, and X 11 are independently deleted, or any naturally occurring amino acid residue except isoleucine, and X 15 are independently a deletion or any naturally occurring amino acid residue, and X 16 are independently a deletion or any naturally occurring amino acid residue except alanine, and X 17 are independently any naturally occurring amino acid residue except valine, and X 21 are independently any naturally occurring amino acid residue except alanine, and X 23are independently any naturally occurring amino acid residue except serine. In some embodiments, X1 comprises tyrosine or glutamic acid; X2 comprises asparagine or aspartic acid; X3 comprises proline or threonine; X4 comprises glutamic acid or asparagine; X5 comprises threonine or asparagine; X6 comprises alanine or proline; X7 comprises alanine or threonine; X8 comprises alanine; X9 comprises tryptophan or arginine; and X 10 Does it contain glycine? X 11 contains tryptophan or serine; X 12 contains aspartic acid, arginine, or serine; X 13 contains arginine, serine, or glycine; X 14 contains tyrosine or glycine or valine; X 15 Does it contain tyrosine? X 16 Does it contain tyrosine? X 17 Does it contain phenylalanine? X 18 contains tyrosine or asparagine; X 19 contains glycine or aspartic acid; X 20 contains threonine or isoleucine; X 21 contains aspartic acid or asparagine; X 22 contains alanine or threonine; X 23 contains asparagine or aspartic acid; X 24 contains isoleucine, serine, or threonine; X 25 contains alanine or threonine, or is a combination thereof.

[0020] In another aspect, the disclosure provides an isolated anti-IBP4 antibody or fragment thereof, comprising: (i) a heavy chain comprising an HCDR1 comprising the amino acid sequence GYTFTDYY (SEQ ID NO: 75), an HCDR2 comprising the amino acid sequence INPNNGGX1 (SEQ ID NO: 360), and an HCDR3 comprising the amino acid sequence ARWX2X3DX4X5FDY (SEQ ID NO: 361); and (ii) an HCDR3 comprising the amino acid sequence QNVGX6X 7( and X7 is independently any naturally occurring amino acid residue except alanine. In some embodiments, X1 comprises alanine or serine; X2 comprises glycine; X3 comprises tryptophan; X4 comprises arginine or glycine; X5 comprises tyrosine or glycine; X6 comprises isoleucine or threonine; X7 comprises aspartic acid or asparagine; X8 comprises serine or isoleucine; X9 comprises alanine or threonine; or a combination thereof.

[0021] In another aspect, the disclosure provides an isolated anti-IBP4 antibody or fragment thereof, comprising: (i) a heavy chain comprising an HCDR1 comprising the amino acid sequence GYTFTDYE (SEQ ID NO: 244), an HCDR2 comprising the amino acid sequence IDX1ETGGX2 (SEQ ID NO: 364), and an HCDR3 comprising the amino acid sequence TRARGSX3SVYYFDX4 (SEQ ID NO: 365); and (ii) a light chain comprising an LCDR1 comprising the amino acid sequence QNVX5TN (SEQ ID NO: 366), an LCDR2 comprising the amino acid sequence SX6S (SEQ ID NO: 367), and an LCDR3 comprising the amino acid sequence QQYDX7YPLT (SEQ ID NO: 368), wherein each of X1, X3, X4, X5, X6, and X7 is independently any naturally occurring amino acid residue, and X2 is independently any naturally occurring amino acid residue except isoleucine. In some embodiments, X1 comprises proline or threonine; X2 comprises proline or alanine; X3 comprises serine or arginine; X4 comprises tyrosine or asparagine; X5 comprises glycine or aspartic acid; X6 comprises alanine or threonine; X7 comprises serine or threonine; or a combination thereof.

[0022] In another aspect, the disclosure provides an isolated anti-IBP4 antibody or fragment thereof, comprising: (i) a heavy chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 68, or a light chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 70; (ii) a heavy chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 103, or a light chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 105. (iii) a heavy chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 209 or a light chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 211; (iv) an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 237. (v) a heavy chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 266 or a light chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 268; (vi) a heavy chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 277. or 99% identity to SEQ ID NO: 279; or (vii) a heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 279; or a light chain comprising an amino acid sequence at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 305; or a light chain comprising an amino acid sequence at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 307.In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 68. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 70. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 68 and a light chain of SEQ ID NO: 70. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 103. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 105. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 103 and a light chain of SEQ ID NO: 105. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 209. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 211. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 209 and a light chain of SEQ ID NO: 211. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 237. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 239. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 237 and a light chain of SEQ ID NO: 239. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 266. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 268. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 266 and a light chain of SEQ ID NO: 268. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 277. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 279. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 277 and a light chain of SEQ ID NO: 279. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 305. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 307. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 305 and a light chain of SEQ ID NO: 307.

[0023] In another aspect, the disclosure provides an isolated anti-IBP4 antibody or fragment thereof, comprising: (i) a heavy chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 123, or a light chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 70; (ii) a heavy chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 128, or a light chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 129; (iii) a heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 333, or a light chain comprising an amino acid sequence at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 211; (iv) a heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 335. (v) a heavy chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 337 or a light chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 268; (vi) a heavy chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 339. , 96%, 97%, 98%, or 99% identity to SEQ ID NO: 279, or a light chain comprising an amino acid sequence at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 279; or (vii) a heavy chain comprising an amino acid sequence at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 341, or a light chain comprising an amino acid sequence at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 307;In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 123. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 70. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 123 and a light chain of SEQ ID NO: 70. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 128. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 105. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 128 and a light chain of SEQ ID NO: 105. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 333. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 211. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 333 and a light chain of SEQ ID NO: 211. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 335. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 239. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 335 and a light chain of SEQ ID NO: 239. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 337. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 268. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 337 and a light chain of SEQ ID NO: 268. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 339. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 279. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 339 and a light chain of SEQ ID NO: 279. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 341. In some embodiments, the antibody or fragment thereof comprises a light chain of SEQ ID NO: 307. In some embodiments, the antibody or fragment thereof comprises a heavy chain of SEQ ID NO: 341 and a light chain of SEQ ID NO: 307.

[0024] In some embodiments, the present disclosure provides an anti-SHBG antibody or fragment thereof, or an anti-IBP4 antibody or fragment thereof, which is an FAB, Fab', F(ab')2, Fv, scFv, (scFv)2, single-chain antibody molecule, dual variable region antibody, single variable region antibody, linear antibody, or V region formed from an antibody fragment. In some embodiments, the antibody or fragment thereof is humanized, partially humanized, chimeric, or humanized chimeric. In some embodiments, the antibody or fragment thereof is conjugated or recombinantly fused to a diagnostic or detectable agent. In some embodiments, the antibody or fragment thereof is bound to a solid support. In some embodiments, the solid support comprises beads.

[0025] In another aspect, the disclosure provides an anti-SHBG or anti-IBP4 antibody or fragment thereof, -7 M or less, 1 x 10 -8 M or less, 1 x 10 -9 M or less, 1 x 10 -10 M or less, 2 x 10 -7 M or less, 2 x 10 -8 M or less, 2 x 10 -9 M or less, 2 x 10 -10 M or less, 5 x 10 -7 M or less, 5 x 10 -8 M or less, 5 x 10 -9 M or less, or 5 x 10 -10 M or less dissociation constant (K D The present invention provides anti-SHBG or anti-IBP4 antibodies or fragments thereof that bind to their respective antigens with an affinity as measured by the affinity (A) of the antibody or fragment thereof.

[0026] In another aspect, the present disclosure provides a nucleic acid molecule encoding the heavy or light chain of an anti-SHBG antibody or fragment thereof, or an anti-IBP4 antibody or fragment thereof. In some embodiments, the nucleic acid molecule encodes the heavy chain of an anti-SHBG antibody or fragment thereof, or an anti-IBP4 antibody or fragment thereof. In some embodiments, the nucleic acid molecule is an isolated nucleic acid molecule. In some embodiments, the nucleic acid molecule encodes the light chain of an anti-SHBG antibody or fragment thereof, or an anti-IBP4 antibody or fragment thereof. In some embodiments, the nucleic acid molecule is an isolated nucleic acid molecule. In some embodiments, the nucleic acid molecule encodes the heavy and light chains of an anti-SHBG antibody or fragment thereof, or an anti-IBP4 antibody or fragment thereof.

[0027] In some embodiments, a nucleic acid molecule encoding a heavy chain has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to at least one of SEQ ID NOs: 3, 35, 200, or 202; or a nucleic acid molecule encoding a light chain has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NOs: 5, 37, 201, or 203. In some embodiments, a nucleic acid molecule encoding a heavy chain has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to at least one of SEQ ID NOs: 67, 102, 204, 206, 208, 236, 265, 276, 304, 334, 336, 338, 340, or 342; or a nucleic acid molecule encoding a light chain has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NOs: 69, 104, 205, 207, 210, 238, 267, 278, or 306. In some embodiments, the disclosure provides a vector comprising the nucleic acid molecule. In some embodiments, the disclosure provides an expression vector comprising the nucleic acid molecule. In some embodiments, the disclosure provides a cell comprising the nucleic acid molecule, vector, or expression vector.

[0028] In another aspect, the present disclosure provides a method for detecting one or more biomarkers in a biological sample, the method comprising: (i) contacting the biological sample containing one or more biomarkers with an anti-SHBG antibody or fragment thereof, or an IBP4 antibody or fragment thereof, and (ii) detecting binding of the antibody or fragment thereof to the one or more biomarkers. In some embodiments, detecting binding comprises a proteomics workflow including mass spectrometry quantification. In some embodiments, the one or more biomarkers comprise SHBG, and the antibody or fragment thereof is an anti-SHBG antibody or fragment thereof. In some embodiments, the one or more biomarkers comprise IBP4, and the antibody or fragment thereof is an anti-IBP4 antibody or fragment thereof. In some embodiments, the one or more biomarkers comprise SHBG and IBP4, and the antibody or fragment thereof comprises an anti-SHBG antibody or fragment thereof and an anti-IBP4 antibody or fragment thereof. In some embodiments, the biological sample comprises blood or serum. In some embodiments, detecting comprises separating the one or more biomarkers bound to the antibody or antibody fragment thereof from the biological sample. In some embodiments, the detecting comprises an immunoassay selected from the group consisting of sandwich immunoassay, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), Western blotting, nephelometry, turbidimetry, immunoradiometric assay, lateral flow, and immunohistochemistry / cytochemistry.

[0029] In another aspect, the present disclosure provides a method for assessing or predicting the health status of a subject, comprising: (i) obtaining a biological sample containing one or more biomarkers from the subject; (ii) contacting an anti-SHBG antibody or a fragment thereof, or an anti-IBP4 antibody or a fragment thereof, with the biological sample under conditions suitable for the antibody or a fragment thereof to bind to the one or more biomarkers; and (iii) detecting binding of the antibody or a fragment thereof to the one or more biomarkers, wherein detecting binding of the antibody to the one or more biomarkers indicates the presence of the one or more biomarkers in the biological sample, and not detecting binding of the antibody to the one or more biomarkers indicates the absence of the one or more biomarkers in the biological sample, and the presence or absence of the one or more biomarkers in the biological sample is used to assess or predict the health status of the subject. In some embodiments, the method further comprises quantifying the amount of one or more biomarkers bound to the antibody or a fragment thereof in the biological sample, and the amount of the one or more biomarkers is used to assess or predict the health status of the subject. In some embodiments, the subject is a pregnant woman. In some embodiments, the health condition comprises any one of abnormal glucocorticoid, gestational diabetes, hypertension, preeclampsia, intrauterine growth restriction, stillbirth, fetal growth restriction, HELLP syndrome, oligohydramnios, chorioamnionitis, chorioamnionitis, placenta previa, placental acreta, abruption, placental abruption, placental hemorrhage, premature premature rupture of membranes, preterm labor, unfavorable cervix, post-term pregnancy, cholelithiasis, or uterine over distention. In some embodiments, detecting comprises isolating one or more biomarkers from the biological sample using an antibody or antibody fragment thereof. In some embodiments, detecting binding comprises a proteomics workflow including mass spectrometry quantification.In some embodiments, detecting comprises an immunoassay selected from the group consisting of sandwich immunoassay, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), Western blotting, nephelometry, nephelometry, immunoradiometry, lateral flow, and immunohistochemistry / cytochemistry. In some embodiments, the one or more biomarkers comprise SHBG, and the antibody or fragment thereof is an anti-SHBG antibody or fragment thereof disclosed herein. In some embodiments, the one or more biomarkers comprise IBP4, and the antibody or fragment thereof is an anti-IBP4 antibody or fragment thereof. In some embodiments, the one or more biomarkers comprise SHBG and IBP4, and the antibody or fragment thereof comprises an anti-SHBG antibody or fragment thereof, an anti-IBP4 antibody or fragment thereof disclosed herein.

[0030] In another aspect, the present disclosure provides a method of treating a subject, the method comprising administering to the subject an effective amount of an anti-SHBG antibody or fragment thereof, or an anti-IBP4 antibody or fragment thereof, as disclosed herein. In some embodiments, the subject is suffering from an SHBG-related disease, disorder, or condition, and the subject is administered an anti-SHBG antibody or fragment thereof. In some embodiments, the SHBG-related disease, disorder, or condition comprises abnormal glucose metabolism, gestational diabetes, hypertension, preeclampsia, intrauterine growth retardation, stillbirth, fetal growth restriction, HELLP syndrome, oligohydramnios, chorioamnionitis, placenta previa, placenta accreta, abruption, placental abruption, placental hemorrhage, premature premature rupture of membranes, preterm labor, adverse cervix, post-term pregnancy, cholelithiasis, placental insufficiency, premature cervical shortening, microbial dysbiosis, or uterine hyperdistension. In some embodiments, the subject is suffering from an IBP4-associated disease, disorder, or condition, and the subject is administered an anti-IBP4 antibody or fragment thereof disclosed herein. In some embodiments, the IBP4-associated disease, disorder, or condition comprises abnormal glucose transporter, gestational diabetes, hypertension, preeclampsia, intrauterine growth retardation, stillbirth, fetal growth restriction, HELLP syndrome, oligohydramnios, chorioamnionitis, chorioamnionitis, placenta previa, placenta accreta, abruption, placental abruption, placental hemorrhage, premature premature rupture of membranes, preterm labor, adverse cervix, post-term pregnancy, cholelithiasis, placental insufficiency, premature cervical shortening, microbial dysbiosis, or uterine hyperdistension.

[0031] In another aspect, the present disclosure provides a composition comprising an anti-SHBG antibody or a fragment thereof, or an anti-IBP4 antibody or a fragment thereof; and a carrier. In some embodiments, the carrier comprises a solid support. In some embodiments, the solid support comprises beads. In some embodiments, the carrier comprises a pharmaceutically acceptable carrier.

[0032] In another aspect, the present disclosure provides a kit comprising an anti-SHBG antibody or fragment thereof, or an anti-IBP4 antibody or fragment thereof; one or more auxiliary reagents; and packaging therefor. In some embodiments, the anti-SHBG antibody or fragment thereof, or the anti-IBP4 antibody or fragment thereof is bound to a solid support. In some embodiments, the solid support comprises beads. In some embodiments, the kit comprises an anti-IBP4 antibody or fragment thereof and an anti-SHBG antibody or fragment thereof. [Brief explanation of the drawings]

[0033] [Figure 1-1] Figures 1A-1B show sequence alignments of the heavy and light chain variable regions of anti-SHBG antibodies designated 4G10.F8 and 13C9.E5. Exemplary complementarity-determining region (CDR) boundaries are in bold. Figure 1A shows a sequence alignment of the heavy chain variable regions of anti-SHBG antibodies 4G10.F8 (SEQ ID NO: 8) and 13C9.E5 (SEQ ID NO: 40). Figure 1B shows a sequence alignment of the light chain variable regions of anti-SHBG antibodies 4G10.F8 (SEQ ID NO: 10) and 13C9.E5 (SEQ ID NO: 42). [Figure 1-2] Same as above.

[0034] [Figure 2-1] Figures 2A-2B show sequence alignments of the heavy and light chain variable regions of anti-IBP4 antibodies designated 5B3.G9 and 7D6.G4. Exemplary CDR boundaries are in bold. Figure 2A shows a sequence alignment of the heavy chain variable regions of anti-IBP4 antibodies 7D6.G4 (SEQ ID NO: 107) and 5B3.G9 (SEQ ID NO: 72). Figure 2B shows a sequence alignment of the light chain variable regions of anti-IBP4 antibodies 5B3.G9 (SEQ ID NO: 74) and 7D6.G4 (SEQ ID NO: 109). [Figure 2-2] Same as above.

[0035] [Figure 3]Figures 3A and 3B show gel images of Western blots demonstrating that the anti-SHBG antibodies described herein bind to human SHBG protein. The gel images show that the anti-SHBG antibodies designated variant 4G10.F8 and variant 13C9.E5 (Figures 3A and 3B, respectively) produce strong signals in human serum containing SHB4 protein. The signals corresponding to the anti-SHBG antibodies are indicated by black arrows.

[0036] [Figure 4] Figure 4 shows Biocore data for exemplary anti-SHBG antibody clones. An exemplary clone, SHBG variant 4G10.F8, is labeled with a black arrow.

[0037] [Figure 5] Figure 5 shows data from a pooled peptide competition assay. The top panel shows a line graph demonstrating that the peptide pool of peptides 1+2+3 competes with variants 4G10.F8 and 13C9.E5 for binding to the target epitope. The y-axis shows the unblocked signal. The x-axis shows the concentration of the antibody assayed. The bottom panel shows a heat map of the data from the line graph in the top panel.

[0038] [Figure 6] Figure 6 shows data from a single peptide competition assay. The top panel shows a line graph of the single peptide competition assay, with the y-axis representing the unblocked signal and the x-axis representing the concentration of antibody assayed. The bottom panel shows a heat map of the data from the competition assay.

[0039] [Figure 7] 7 shows the amino acid sequence of the SHBG target epitope mapped to blocking peptide 1+2+3 (SEQ ID NO: 120). The shaded amino acids indicate the signal sequence not present in the mature SHBG protein. Peptides 1+2+3 are mapped to the underlined portion of the amino acid sequence.

[0040] [Figure 8] Figures 8A and 8B show data from an electrochemiluminescence immunoassay (ECLIA) characterizing the effect of SHBG glycosylation on antibody binding. Figure 8A shows the percent remaining signal in an ECLIA assay using anti-SHBG antibody variant 13C9.E5 to bind to SHBG serum from pregnant and non-pregnant women. Figure 8B shows that samples treated with deglycosylation enzymes (wells labeled "+") produce a lower signal than cells not treated with deglycosylation enzymes (e.g., compare wells 1 and 2).

[0041] [Figure 9] 9 shows the amino acid sequence of the SHBG epitope (SEQ ID NO: 121). The glycosylation site of the SHBG epitope is labeled with a black arrow at position T36. An additional site for glycosylation is identified by an underlined "N." The underlined portion of the epitope is recognized by the SHBG variant antibodies 4G10.F8 and 13C9.E5.

[0042] [Figure 10] Figure 10 shows a gel image of a Western blot confirming that anti-IBP4 antibody clone variant 5B3.G9 and variant 7D6.G4 bind to human IBP4 protein. Signals corresponding to anti-IBP4 variant antibodies 5B3.G9 and 7D6.G4 are indicated by black arrows. Additional candidate antibodies assayed are indicated above their respective lanes.

[0043] [Figure 11]Figures 11A and 11B show data from a direct competitive binding assay, including an electrochemiluminescence immunoassay (ECLIA), performed using anti-IBP4 variant antibody clones 5B3.G9 and 7D6.G4. Figure 11A shows that the signal from clone variant 5B3.G9 decreases with increasing concentrations of variant 7D6.G4. Figure 11B shows that the signal from variant 7D6.G4 decreases with increasing concentrations of antibody clone variant 5B3.G9.

[0044] [Figure 12] FIG. 12 shows data from an ECLIA competitive binding assay mapping the epitopes of the variant 5B3.G9 and variant 7D6.G4 antibody clones (SEQ ID NOs: 344-353, 369).

[0045] [Figure 13] Figures 13A-13B show correlation plots of IBP4 response ratios (RR) measured by alternating current mass spectrometry (AC-MS) and an antibody enrichment assay referred to herein as CLIA. Figure 13A shows data from beads coupled to the IBP4-5B3.G9 antibody for AC-MS. Figure 13B shows beads coupled to the IBP4-7D6.G4 antibody for AC-MS. The IBP4 response ratios from each AC-MS assay were plotted against the IBP4 RR data from the CLIA assay.

[0046] [Figure 14-1]Figures 14A-14F show correlation plots of anti-SHBG antibody response ratios (RR) from the AC-MS assay and the antibody enrichment assay referred to as CLIA. Data from assays using beads coupled to anti-SHBG antibodies are shown in Figure 14A: variant 13C9.E5; Figure 14B: 4G10.F8; Figure 14C: 16A6.A2; Figure 14D: 1G7.C5; Figure 14E: 11C6.A8; and Figure 14F: SHBG-6001 control anti-SHBG antibody. The SHBG response ratios from the AC-MS assay were plotted against the response ratios from the CLIA assay. [Figure 14-2] Same as above.

[0047] [Figure 15-1] FIG. 15 shows AC-MS data from a combination of IBP4 and SHBG beads. [Figure 15-2] Same as above.

[0048] [Figure 16] Figure 16 shows direct ECLIA data generated for five additional anti-IBP4 antibodies (1H9.B6, 5C11.A7, 7G11.H4, 3D4.E9, and 1G3.A3) compared to 5B3.G9, demonstrating their similar affinity with 5B3.G9 as a reference.

[0049] [Figure 17] FIG. 17 shows competition direct ECLIA data demonstrating the ability of each of the anti-IBP4 antibodies to block 5B3.G9 from binding to its epitope, thus suggesting that they bind to the same or similar epitopes.

[0050] [Figure 18-1]Figures 18A-18C show correlation plots comparing AC-MS and CLIA IBP4 response ratios (RR). Figure 18A shows M-PVA beads coupled to the 1H9.B6 anti-IBP4 antibody for AC-MS. Figure 18B shows beads coupled to the 5C11.A7 anti-IBP4 antibody for AC-MS. Figure 18C shows beads coupled to the 7G11.H4 anti-IBP4 antibody for AC-MS. The IBP4 response ratios from each AC-MS assay were plotted against the IBP4 RR data from the CLIA assay. [Figure 18-2] Same as above. [Figure 18-3] Same as above.

[0051] [Figure 19-1] Figure 19 shows the heavy chain CLUSTAL O(1.2.4) full-length multiple sequence alignment for the 1G3.A3 (sequence number 305), 1H9.B6 (sequence number 209), 3D4.E9 (sequence number 277), 5C11.A7 (sequence number 237), 7G11.H4 (sequence number 266), 7D6.G4 (sequence number 103), and 5B3.G9 (sequence number 68) anti-IBP4 antibodies. [Figure 19-2] Same as above.

[0052] [Figure 20] Figure 20 shows the light chain CLUSTAL O(1.2.4) full-length multiple sequence alignment for 1G3.A3 (sequence number 307), 1H9.B6 (sequence number 70), 3D4.E9 (sequence number 279), 5C11.A7 (sequence number 239), 7G11.H4 (sequence number 268), 7D6.G4 (sequence number 105), and 5B3.G9 (sequence number 70) anti-IBP4 antibodies.

[0053] [Figure 21-1]Figures 21A-21B show the heavy and light chain IMGT consensus CDRs (complementarity-determining regions) for anti-IBP4 antibodies 1H9.B6, 5C11.A7, 7G11.H4, 5B3.G9, 7D6.G4, 3D4.E9, and 1G3.A3. The CDR nomenclature is shown below the sequences. Figure 21A shows the heavy chain IMGT consensus CDRs. The sequences for each CDRH1, CDRH2, and CDRH3 are shown in the left, center, and right columns, respectively (SEQ ID NOs: 275, 216, and 217 for 1H9.B6; SEQ ID NOs: 244, 245, and 246 for 5C11.A7 and 7G11.H4; SEQ ID NOs: 75, 216, and 77 for 5B3.G9 and 7D6.G4; SEQ ID NOs: 244, 284, and 285 for 3D4.E9; and SEQ ID NOs: 244, 312, and 313 for 1G3.A3). Figure 21B shows the light chain IMGT consensus CDRs. The sequences for each CDRL1, CDRL2, and CDRL3 are shown in the left, center, and right columns, respectively (sequence numbers 226, 24, and 227 for 1H9.B6; sequence numbers 87, 24, and 259 for 5C11.A7; sequence numbers 23, 24, and 259 for 7G11.H4; sequence numbers 87, 24, and 89 for 5B3.G9; sequence numbers 87, 24, and 99 for 7D6.G4; sequence numbers 87, 297, and 298 for 3D4.E9; and sequence numbers 323, 24, and 324 for 1G3.A3). [Figure 21-2] Same as above.

[0054] [Figure 22-1]Figures 22A-22B show the heavy and light chain variable regions for the anti-IBP4 antibodies 1G3.A3, 1H9.B6, 3D4.E9, 5C11.A7, 7G11.H4, 7D6.G4, and 5B3.G9. Bold and underlined residues indicate less favored residues from the 1G3.A3 clone according to the alignment of these antibodies. Figure 22A shows the heavy chain variable regions for 1G3.A3 (SEQ ID NO: 309), 1H9.B6 (SEQ ID NO: 213), 3D4.E9 (SEQ ID NO: 281), 5C11.A7 (SEQ ID NO: 241), 7G11.H4 (SEQ ID NO: 241), 7D6.G4 (SEQ ID NO: 107), and 5B3.G9 (SEQ ID NO: 72). Figure 22B shows the light chain variable regions for 1G3.A3 (sequence number 311), 1H9.B6 (sequence number 215), 3D4.E9 (sequence number 283), 5C11.A7 (sequence number 243), 7G11.H4 (sequence number 271), 7D6.G4 (sequence number 109), and 5B3.G9 (sequence number 74). [Figure 22-2] Same as above.

[0055] [Figure 23-1]Figures 23A-23B show CLUSTAL O(1.2.4) multiple sequence alignments of the heavy chain variable regions of two groups of anti-IBP4 antibodies: 1H9.B6, 7D6.G4, 5B3.G9, and 1G3.A3 (Group 1) in one group, and 3D4.E9, 5C11.A7, 7G11.H4, and 1G3.A3 (Group 2) in the other group. The HCDRs for each alignment and group are highlighted and are the same as those in Figures 21A-21B. Bold and underlined residues indicate less favored residues from the 1G3.A3 clone according to the alignment of these antibodies. The CDR nomenclature is shown below the sequence alignment for each group. Figure 23A shows a sequence alignment for the heavy chain variable regions of group 1: 1G3.A3 (SEQ ID NO: 309), 1H9.B6 (SEQ ID NO: 213), 7D6.G4 (SEQ ID NO: 107), and 5B3.G9 (SEQ ID NO: 72). Figure 23B shows a sequence alignment for the heavy chain variable regions of group 2: 1G3.A3 (SEQ ID NO: 309), 3D4.E9 (SEQ ID NO: 281), 5C11.A7 (SEQ ID NO: 241), and 7G11.H4 (SEQ ID NO: 241). [Figure 23-2] Same as above.

[0056] [Figure 24-1]Figures 24A-24B show CLUSTAL O(1.2.4) multiple sequence alignments of the light chain variable regions of two groups of anti-IBP4 antibodies: 1H9.B6, 7D6.G4, 5B3.G9, and 1G3.A3 (Group 1) in one group, and 3D4.E9, 5C11.A7, 7G11.H4, and 1G3.A3 (Group 2) in the other group. The LCDRs for each alignment and group are highlighted and are the same as those in Figures 21A-21B. Bold and underlined residues indicate less favored residues from the 1G3.A3 clone according to the alignment of these antibodies. The CDR nomenclature is shown below the sequence alignment for each group. Figure 24A shows a sequence alignment for the light chain variable regions of group 1: 1G3.A3 (SEQ ID NO: 311), 1H9.B6 (SEQ ID NO: 215), 7D6.G4 (SEQ ID NO: 109), and 5B3.G9 (SEQ ID NO: 74). Figure 24B shows a sequence alignment for the light chain variable regions of group 2: 1G3.A3 (SEQ ID NO: 311), 3D4.E9 (SEQ ID NO: 283), 5C11.A7 (SEQ ID NO: 243), and 7G11.H4 (SEQ ID NO: 271). [Figure 24-2] Same as above. DETAILED DESCRIPTION OF THE INVENTION

[0057] Detailed Description of the Invention Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is for the purpose of providing a context for aspects of the present disclosure. Such discussion is not an admission that any or all of these matters form part of the prior art with respect to any embodiment disclosed herein.

[0058] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. Unless otherwise defined, certain terms used herein have the meanings set forth herein.

[0059] The present disclosure provides SHBG and IBP4 binding agents. Such agents include antibodies or antigen-binding fragments thereof that bind to SHBG or IBP4. Such binding agents are useful in compositions and various methods for detecting the presence (including quantifying the amount) or absence of SHBG and / or IBP4 in a sample, including methods for determining the probability of preterm birth and preeclampsia. The binding agents described herein are also useful for treating certain health conditions. definition

[0060] The terms "SHBG," "sex hormone binding protein," "SHBG polypeptide," and similar terms, unless otherwise indicated, refer to a polypeptide ("polypeptide" and "protein" are used interchangeably herein) or any native SHBG from any vertebrate source, including mammals such as primates (e.g., humans, cynomolgus monkeys (cynos)), dogs, and rodents (e.g., mice and rats). The term SHBG includes any species homolog. The term SHBG also includes any SHBG that is naturally expressed by cells or that can be expressed in cells transfected with a gene or cDNA encoding the polypeptide. SHBG, also known in the art as androgen-binding protein (ABG) and testosterone-estrogen-binding globulin (TEBG), has an N-terminal steroid-binding and dimerization domain and a C-terminal domain containing a highly conserved consensus sequence for glycosylation.

[0061] SHBG is a steroid-binding protein secreted by the liver that functions as a regulator of steroid response. SHBG is secreted into the blood by the liver and can bind to androgens and estrogens. Furthermore, in pregnant women, SHBG has been reported in the human placenta. Beyond regulating steroid response, SHBG plays a role in predicting preterm birth, prolonged neonatal hospital stay, and increased severity of adverse neonatal outcomes. See Markenson et al., "Performance of a proteomic preterm delivery predictor in a large independent prospective cohort." American Journal of Obstetrics & Gynecology, 2(3): 100140 (2020).

[0062] The term SHBG encompasses not only "full-length" unprocessed SHBG, but also any form of SHBG or any fragment thereof resulting from processing in cells, including the four known alternatively spliced ​​isoforms of SHBG that differ in the length of their intracellular tails. The term SHBG also encompasses naturally occurring variants of SHBG, such as SNP variants, splice variants, and allelic variants. Unless otherwise noted, preferably, SHBG is human SHBG. The full-length human SHBG amino acid sequence is found in GenBank Accession No. NP_001031:MESRGPLATSRLLLLLLLLLLRHTRQGWALRPVLPTQSAHDPPAVHLSNGPGQEPIAVMTFDLTKITKTSSSFEVRTWDPEGVIFYGDTNPKDDWFMLGLRDGRPEIQLHNHWAQLTVGAGPRLDDGRWHQVEVKMEGDSVLLEVDGEEVLRLRQVSGPLTSKRHPIMRIALGGLL FPASNLRLPLVPALDGCLRRDSWLDKQAEISASAPTSLRSCDVESNPGIFLPPGTQAEFNLRDIPQPHAEPWAFSLDLGLKQAAGSGHLLALGTPENPSWLSLHLQDQKVVLSSGSGPGLDLPLVLGLPLQLKLSMSRVVLSQGSKMKALALPPLGLAPLLNLWAKPQGRLFLGALPGEDSSTSFCLNGLWAQGQRLDVDQALNRSHEIWTHSC PQSPGNGTDASH (SEQ ID NO: 1).

[0063] Other related SHBG polypeptides encompassed by the term SHBG include fragments, derivatives (e.g., substitution, deletion, truncation, and insertion variants), fusion polypeptides, and interspecies homologs that retain SHBG activity. As will be understood by those skilled in the art, the SHBG binding agents (e.g., antibodies) described herein can bind to SHBG polypeptides, SHBG polypeptide fragments, SHBG antigens, and / or SHBG epitopes. An epitope may be part of a larger SHBG antigen, which may be part of a larger SHBG polypeptide fragment, which may in turn be part of a larger SHBG polypeptide. SHBG may exist in a native or denatured form. The SHBG polypeptides described herein may be isolated from various sources, for example, from human tissue types or from other sources, or may be prepared by recombinant or synthetic methods. SHBG polypeptides may include polypeptides having the same amino acid sequence as the corresponding naturally occurring SHBG polypeptide. Orthologs of SHBG polypeptides are also well known in the art.

[0064] The terms "IBP4," "insulin-like growth factor binding protein 4," "IBP4 polypeptide," and similar terms, unless otherwise indicated, refer to the polypeptide or any native IBP4 from any vertebrate source, including mammals such as primates (e.g., humans, cynomolgus monkeys (cynos)), dogs, and rodents (e.g., mice and rats). The term IBP4 also includes any species homolog. The term IBP4 also includes any IBP4 that is naturally expressed by a cell or that can be expressed in a cell transfected with a gene or cDNA encoding the polypeptide. IBP4, also known in the art as "insulin-like growth factor binding protein 4" (IGFBP4), has three structurally distinct domains that make up one-third of the molecule, with domain 1 being the IGF-binding domain.

[0065] IBP4 is a binding protein secreted by the ovaries, uterus, liver, and placenta that extends the half-life of insulin-like growth factors. Beyond regulating growth responses, IBP4 plays a role in predicting preterm birth, prolonged neonatal hospital stay, and increased severity of adverse neonatal outcomes. See Markenson et al., "Performance of a proteomic preterm delivery predictor in a large independent prospective cohort." American Journal of Obstetrics & Gynecology, 2(3): 100140 (2020).

[0066] The term IBP4 encompasses not only "full-length," unprocessed IBP4, but also any form of IBP4 or any fragment thereof that results from processing in cells, including the four known alternatively spliced ​​isoforms of IBP4 that differ in the length of their intracellular tails. The term IBP4 also encompasses naturally occurring variants of IBP4, such as SNP variants, splice variants, and allelic variants. Unless otherwise specified, preferably, IBP4 is human IBP4. The full-length human IBP4 amino acid sequence is provided by GenBank accession number NP_001543:MLPLCLVAALLLAAGPGPSLGDEAIHCPPCSEEKLARCRPPVGCEELVREPGCGCCATCALGLGMPCGVYTPRCGSGLRCYPPRGVEKPLHTLMHGQGVCMELAEIEAIQESLQPSDKDEGDHPNNSFSPCSAHDRRCLQKHFAKIRDRSTSGGKMKVNGAPREDARPVPQGSCQSELHRALERLAASQSRTHEDLYIIPIPNCDRNGNFHPKQCHPALDGQRGKCWCVDRKTGVKLPGGLEPKGELDCHQLADSFRE (SEQ ID NO: 2).

[0067] Other related IBP4 polypeptides encompassed by the term IBP4 include fragments, derivatives (e.g., substitution, deletion, truncation, and insertion variants), fusion polypeptides, and interspecies homologs that retain IBP4 activity. As will be understood by those skilled in the art, the IBP4-binding agents (e.g., antibodies) described herein can bind to IBP4 polypeptides, IBP4 polypeptide fragments, IBP4 antigens, and / or IBP4 epitopes. An epitope may be part of a larger IBP4 antigen, which may be part of a larger IBP4 polypeptide fragment, which may in turn be part of a larger IBP4 polypeptide. IBP4 may exist in native or denatured forms. The IBP4 polypeptides described herein may be isolated from various sources, for example, from human tissue types or from another source, or may be prepared by recombinant or synthetic methods. IBP4 polypeptides may include polypeptides having the same amino acid sequence as the corresponding naturally occurring IBP4 polypeptide. Orthologs of IBP4 polypeptides are also well known in the art.

[0068] The terms "treating," "treatment," and the like are used herein to generally mean obtaining a desired pharmacological and / or physiological effect. The effect may be prophylactic, in terms of completely or partially preventing a disease, and / or therapeutic, in terms of partially or completely curing a disease and / or adverse effects resulting from a disease. Treatment, as used herein, encompasses any intervention in a subject, including preventing a disease from occurring in a subject who has not yet been diagnosed with it but may be predisposed to the disease, inhibiting a disease, i.e., halting its development, or alleviating a disease, i.e., causing regression of the disease. Treatment also includes any approach for obtaining beneficial or desired results, including clinical results. Non-limiting beneficial or desired results may include one or more of the following: reduction or amelioration of one or more symptoms, reduction in the severity of a condition (including a disease), stabilization (i.e., not worsening) of the status of a condition (including a disease), or delay or slowing of a condition (including a disease).

[0069] It should be noted that as used herein, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise, and are used interchangeably with "at least one" and "one or more." Thus, for example, reference to "a binding agent" includes a mixture of two or more binding agents, and the like.

[0070] Unless otherwise stated, any numerical value, e.g., a concentration or concentration range described herein, should be understood to be modified in all instances by the term "about." Thus, numerical values ​​typically include ±10% of the recited value. For example, a concentration of 1 mg / mL includes 0.9 mg / mL to 1.1 mg / mL. Similarly, a concentration range of 1% to 10% (w / v) includes 0.9% (w / v) to 11% (w / v). As used herein, the use of numerical ranges expressly includes all possible subranges within that range, all individual numerical values, and, unless the context clearly dictates otherwise, includes integers within such ranges and fractions of said values.

[0071] Unless otherwise indicated, the term "at least" preceding a series of elements should be understood to refer to every element in the series. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the present disclosure. Such equivalents are intended to be encompassed by the present disclosure.

[0072] As used herein, the term "binding agent" or its grammatical equivalents refers to a molecule (e.g., an antibody) that has one or more antigen-binding sites that bind to an antigen.For example, the SHBG binding agent described herein includes an antibody, an antibody fragment, or other peptide-based molecule that binds to SHBG.As another example, the IBP4 binding agent described herein includes an antibody, an antibody fragment, or other peptide-based molecule that binds to IBP4.

[0073] As used herein, the term "antibody" is used broadly and includes any "immunoglobulin" or "Ig" molecule, specifically human, humanized, composite, and chimeric antibodies, whether polyclonal or monoclonal, as well as antibody fragments (including agonists, antagonists, neutralizing antibodies, and full-length monoclonal antibodies), antibody compositions with polyepitopic or monoepitopic specificity, recombinantly produced antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), synthetic antibodies, chimeric antibodies, humanized antibodies, or human versions of antibodies with full-length heavy and / or light chains. Generally, antibodies are proteins or peptide chains that exhibit binding specificity to a specific antigen. Antibodies may be agonistic or antagonistic. The structure of antibodies is well known.

[0074] The present disclosure also encompasses antibody fragments (and / or polypeptides comprising antibody fragments) that retain SHBG or IBP4-binding properties. Non-limiting examples of antibody fragments include antigen-binding and / or effector regions of antibodies, such as Fab, Fab', F(ab')2, Fv, scFv, (scFv)2, single-chain antibody molecules, dual-variable-region antibodies, single-variable-region antibodies, linear antibodies, V regions, multispecific antibodies formed from antibody fragments, F(ab)2, Fd, Fc, diabodies, di-diabodies, disulfide-linked Fvs (dsFv), single-domain antibodies (e.g., nanobodies), or other fragments (e.g., fragments consisting of non-covalently coupled heavy and light chain variable regions). In general terms, the variable (V) region domain can be any suitable arrangement of immunoglobulin heavy (VH) and / or light (VL) chain variable domains. For example, the present disclosure also includes tetrameric antibodies comprising two heavy chain molecules and two light chain molecules, each of which is an antibody light chain monomer and an antibody heavy chain monomer. Thus, for example, the V region domain may be dimeric and contain a VH-VH, VH-VL, or VL-VL dimer that binds to SHBG or IBP4. If desired, the VH and VL chains may be covalently coupled, either directly or through a linker, to form a single-chain Fv (scFv). For ease of reference, scFv proteins are referred to herein as being included in the category of "antibody fragments." Another form of antibody fragment is a peptide comprising one or more complementarity-determining regions (CDRs) of an antibody. CDRs (also called "minimal recognition units" or "hypervariable regions") can be obtained by constructing polynucleotides encoding the CDRs of interest.Such polynucleotides are prepared, for example, by synthesizing the variable region using the polymerase chain reaction, using mRNA from antibody-producing cells as a template (see, e.g., Larrick et al., Methods: A Companion to Methods in Enzymology, 2:106 (1991); Courtenay-Luck, "Genetic Manipulation of Monoclonal Antibodies," in Monoclonal Antibodies Production, Engineering and Clinical Application, Ritter et al. (eds.), page 166, Cambridge University Press (1995); and Ward et al., "Genetic Manipulation and Expression of Antibodies," in Monoclonal Antibodies: Principles and Applications, Birch et al., (eds.), page 137, Wiley-Liss, Inc. (1995)). Antibody fragments may be incorporated into single-domain antibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, variable domains of new antigen receptors (v-NARs), and bis-single-chain Fv regions (see, e.g., Hollinger and Hudson, Nature Biotechnology, 23(9):1126-1136, 2005). The binding agent, in some embodiments, contains one or more constant regions, including light and / or heavy chain constant regions, e.g., one or more IgG1, IgG2, IgG3, and / or IgG4 constant regions. In some embodiments, the antibody may comprise an epitope-binding fragment of any of the above. The antibodies described herein may be of any class (e.g., IgG, IgE, IgM, IgD, and IgA) or any subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) of immunoglobulin molecules.IgA and IgG are further subdivided into isotypes IgA1, IgA2, IgG1, IgG2, IgG3, and IgG4. Antibody light chains of vertebrate species can be assigned to one of two distinct types, kappa and lambda, based on the amino acid sequence of their constant domains. Thus, antibodies of the present disclosure may contain kappa or lambda light chain constant domains. According to certain embodiments, the antibodies disclosed herein comprise heavy and / or light chain constant regions derived from murine or human antibodies. In addition to the heavy and light constant domains, antibodies contain an antigen-binding region composed of a light chain variable region and a heavy chain variable region, each of which contains three domains (i.e., complementarity-determining regions 1-3; CDR1, CDR2, and CDR3). The light chain variable region domains are alternatively referred to as LCDR1, LCDR2, and LCDR3, and the heavy chain variable region domains are alternatively referred to as HCDR1, HCDR2, and HCDR3.

[0075] As used herein, the term "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigen specificities (e.g., an isolated antibody that specifically binds to SHBG is substantially free of antibodies that do not bind to SHBG). In addition, an isolated antibody is substantially free of other cellular material and / or chemicals.

[0076] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., antibodies that are identical except for possible naturally occurring mutations that may be present in minor amounts. The monoclonal antibodies disclosed herein can be produced by hybridoma methods, phage display technology, single lymphocyte gene cloning technology, or recombinant DNA methods. For example, monoclonal antibodies can be produced by hybridomas containing B cells obtained from a transgenic non-human animal, such as a transgenic mouse or rat, whose genome contains human heavy chain transgenes and light chain transgenes.

[0077] As used herein, the term "humanized antibody" refers to a non-human antibody that has been modified to increase its sequence homology to that of a human antibody, such that the antigen-binding properties of the antibody are retained but its antigenicity in the human body is reduced.

[0078] As used herein, an antibody that "specifically binds to SHBG" or "specifically binds to IBP4" is an antibody that specifically binds to SHBG or IBP4, preferably human SHBG or IBP4, at a concentration of 1×10 -7 M or less, preferably 1 x 10 -8 M or less, more preferably 5 x 10 -9 M or less, 1 x 10 -9 M or less, 5 x 10 -10 M or less, or 1 x 10 -10 M or less K D For example, the anti-SHBG antibodies described herein bind at about 1.0 x 10 -9 M ~ approx. 2.5×10 -9 K of M D The anti-IBP4 antibodies described herein may have a titer of about 3×10 -8 M ~ approx. 8×10 -8 K of M D The term "K D " refers to the dissociation constant, which is K d K a to (i.e., K d / K a ) and expressed as molar concentration (M). D Values ​​can be determined using methods in the art in light of the present disclosure. For example, the K D can be determined by using surface plasmon resonance, for example, by using a biosensor system, such as a Biacore® system, or by using biolayer interference technology, for example, an Octet RED96 system. D The smaller the value of , the higher the affinity with which the antibody binds to the target antigen.

[0079] The terms "identical" or percent "identity," in the context of two or more nucleic acid or polypeptide sequences (e.g., anti-SHBG antibodies and the polynucleotides encoding them, SHBG polypeptides and the SHBG polynucleotides encoding them, anti-IBP4 antibodies and the polynucleotides encoding them, IBP4 polypeptides and the IBP4 polynucleotides encoding them), refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same when compared and aligned for maximum correspondence as determined using one of the following sequence comparison algorithms or by visual inspection:

[0080] For sequence comparison, typically, one sequence serves as the reference sequence with which test sequence is compared.When using sequence comparison algorithm, test sequence and reference sequence are input into computer, and then coordinates are designated as necessary, and parameter of sequence algorithm program is designated.Then, sequence comparison algorithm calculates the sequence identity percentage of test sequence and reference sequence according to designated program parameter.

[0081] Optimal alignment of sequences for comparison can be determined, for example, by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computer implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by visual inspection (generally, Current Protocols in Molecular Biology, F.M. Ausubel et al., eds., Current Protocols, a joint venture between Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., (1995 Supplement) (Ausubel)).

[0082] Examples of algorithms suitable for determining percent sequence identity and percent sequence similarity are the BLAST and BLAST 2.0 algorithms described in Altschul et al. (1990) J. Mol. Biol. 215: 403-410 and Altschul et al. (1997) Nucleic Acids Res. 25: 3389-3402, respectively. Software for performing BLAST analysis is publicly available through the National Center for Biotechnology Information. This algorithm involves first identifying high-scoring sequence pairs (HSPs) by identifying short words of length W in a query sequence that, when aligned with words of the same length in database sequences, match or meet some positive threshold score T. T is called the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits serve as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased.

[0083] Cumulative scores are calculated using the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0) for nucleotide sequences. For amino acid sequences, a scoring matrix is ​​used to calculate the cumulative score. Extension of word hits in each direction is stopped when the cumulative alignment score deviates from its maximum achieved value by an amount X, when the cumulative score becomes zero or less due to the accumulation of one or more negative-scoring residue alignments, or when the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a word length (W) of 11, an expectation (E) of 10, M=5, N=-4, and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a word length (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915 (1989)).

[0084] In addition to calculating percent sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, for example, Karlin & Altschul, Proc. Nat'l. Acad. Sci. USA 90:5873-5787 (1993)). One measure of similarity provided by the BLAST algorithm is the minimum sum probability (P(N)), which provides an indication of the probability that a match between two nucleotide or amino acid sequences will occur by chance. For example, a nucleic acid is considered to be similar to a reference sequence if the minimum sum probability in the comparison of the test nucleic acid with the reference nucleic acid is less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001.

[0085] Another indicator that two nucleic acid sequences or polypeptides are substantially identical is that the polypeptide encoded by the first nucleic acid is immunologically cross-reactive with the polypeptide encoded by the second nucleic acid, as described below.Therefore, a polypeptide is typically substantially identical to a second polypeptide, for example, when the two peptides differ only by conservative substitutions.Another indicator that two nucleic acid sequences are substantially identical is that the two molecules hybridize with each other under stringent conditions.

[0086] As used herein, the term "label" refers to a molecular entity that emits a signal and can be used as a readout or measurement for the detection of an analyte. Various classes of labels exist. Such labels include fluorophores, enzymes, chemiluminescent moieties, radioactive moieties, organic dyes, small molecules, polypeptides, or functional fragments thereof. Examples of fluorophores include fluorescent dyes such as phycoerythrin (PE), fluorescein isothiocyanate (FITC), tetramethylrhodamine (TRITC), BODIPY, and AlexaFluor® dyes. Fluorescent dyes may also include fluorescence resonance energy transfer (FRET) dyes or time-resolved (TR)-FRET dyes. Fluorophore labels also include fluorescent proteins, such as green fluorescent protein (GFP) and cyan fluorescent protein (CFP). Examples of enzyme labels include alkaline phosphatase (AP) or horseradish peroxidase (HRP). When the substrates 3,3',5,5'-tetramethylbenzidine (TMB), 3,3'-diaminobenzidene (DAB), or 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) are applied to HRP, a colored (chromogenic) or light (chemiluminescent) signal is generated. Radioactive moiety labels include carbon-14 or tritium. Small molecule labels include biotin, resins such as agarose beads and fluorescently labeled magnetic beads, or nanoparticles such as colloidal gold. Polypeptide or functional fragment labels include avidin, streptavidin, or neutravidin, which have affinity for biotin. Polypeptide or functional fragment labels also include hemagglutinin (HA), glutathione-S-transferase (GST), or c-myc.

[0087] The label of the present disclosure can be conjugated to any of the antibodies, antigen-binding fragments thereof, or binding agents described herein. Conjugation can include non-covalent or covalent crosslinking as described above. In some configurations, labels conjugated to detection probes require an additional substrate or binding agent as described above. For example, an HRP label conjugated to an anti-SHBG antibody or its antigen-binding fragment requires the substrate disclosed above. Many other labeling configurations are known in the art. The present disclosure includes all labeling configurations exemplified herein and / or known in the art. In some embodiments, the labeling configuration can include PE conjugated to an anti-SHBG antibody, antigen-binding fragment thereof, or binding agent.

[0088] As used herein, the term "preterm birth" refers to delivery or birth at less than 37 completed weeks of gestation. Other commonly used subcategories of preterm birth have been established and are defined as slightly preterm (delivery at 33-36 weeks of gestation), very preterm (delivery at <33 weeks of gestation), and extremely preterm (delivery at ≤28 weeks of gestation). With respect to the methods disclosed herein, those skilled in the art will understand that the cutoffs defining preterm birth from full-term birth, as well as the cutoffs defining subcategories of preterm birth, can be adjusted during the practice of the methods disclosed herein to, for example, maximize certain health benefits. In various embodiments of the present disclosure, cutoffs defining preterm birth include, for example, birth at ≤ 37 weeks gestation, birth at ≤ 36 weeks gestation, birth at ≤ 35 weeks gestation, birth at ≤ 34 weeks gestation, birth at ≤ 33 weeks gestation, birth at ≤ 32 weeks gestation, birth at ≤ 30 weeks gestation, birth at ≤ 29 weeks gestation, birth at ≤ 28 weeks gestation, birth at ≤ 27 weeks gestation, birth at ≤ 26 weeks gestation, birth at ≤ 25 weeks gestation, birth at ≤ 24 weeks gestation, birth at ≤ 23 weeks gestation, or birth at ≤ 22 weeks gestation. In some embodiments, the cutoff defining preterm birth is ≤ 35 weeks gestation. It is further understood that such adjustments are well within the skill set of individuals considered skilled in the art and are encompassed within the scope of the disclosure. Gestational age is a surrogate indicator of fetal development and fetal readiness. Gestational age is typically defined as the time from the date of the last normal menstrual period to the date of delivery. However, obstetric measures and ultrasound estimates can also assist in estimating gestational age. Preterm births are generally classified into two distinct subgroups: spontaneous preterm birth, which occurs after the spontaneous onset of preterm labor or preterm rupture of membranes before term, regardless of subsequent augmentation of labor or cesarean section; and medically indicated preterm birth, which occurs after induction or cesarean section and in the absence of spontaneous onset of labor due to one or more conditions that the female caregiver determines threaten the health or life of the mother and / or fetus. Voluntary preterm births for reasons other than life-threatening may also be referred to as medically indicated.In some embodiments, the methods disclosed herein are directed to determining the probability of spontaneous preterm birth or medically indicated preterm birth. In some embodiments, the methods disclosed herein are directed to determining the probability of spontaneous preterm birth. In additional embodiments, the methods disclosed herein are directed to medically indicated preterm birth. In additional embodiments, the methods disclosed herein are directed to predicting gestational age at birth.

[0089] As used herein, the term "biological sample" encompasses any sample collected from a pregnant woman and contains one or more of the biomarkers disclosed herein. Suitable samples in the context of this disclosure include, for example, blood, plasma, serum, amniotic fluid, vaginal secretions, saliva, and urine. In some embodiments, the biological sample is selected from the group consisting of whole blood, plasma, and serum. In certain embodiments, the biological sample is serum. As will be understood by those skilled in the art, a biological sample may include any fraction or component of blood, including, but not limited to, T cells, monocytes, neutrophils, red blood cells, platelets, and microvesicles, such as exosomes and exosome-like vesicles. In certain embodiments, the biological sample is serum.

[0090] As used herein, the term "diagnostic agent" or "detectable agent" refers to a compound that can be used to detect, purify, isolate, or enrich a target, particularly a biomarker.

[0091] A "proteomics workflow" generally involves one or more of the following steps: thawing a serum sample and depleting the 14 most abundant proteins using immunoaffinity chromatography. The depleted serum is digested with a protease, such as trypsin, to generate peptides. The digest is then enriched with a mixture of SIS peptides, desalted, and subjected to LC-MS / MS with a triple quadrupole instrument operated in MRM mode. A response ratio is generated from the area ratio of the endogenous peptide peak and the corresponding SIS peptide equivalent peak. Those skilled in the art will recognize that other types of MS, such as MALDI-TOF or ESI-TOF, can be used in the disclosed methods. Additionally, those skilled in the art can modify the proteomics workflow by selecting specific reagents (such as proteases) or by omitting or changing the order of certain steps. For example, immunodepletion may not be necessary, SIS peptides may be added earlier or later, or stable isotope-labeled proteins may be used as standards instead of peptides.

[0092] As used herein, the term "mass spectrometer" refers to a device capable of volatilizing / ionizing analytes, forming gas-phase ions, and determining their absolute or relative molecular mass. Suitable methods of volatilization / ionization are matrix-assisted laser desorption / ionization (MALDI), electrospray, laser / light, thermal, electrical, atomization / nebulization, etc., or combinations thereof. Suitable forms of mass spectrometry include, but are not limited to, ion trap instruments, quadrupole instruments, electrostatic sector instruments, time-of-flight instruments, time-of-flight tandem mass spectrometers (TOF MS / MS), Fourier transform mass spectrometers, Orbitrap, and hybrid instruments composed of various combinations of these types of mass analyzers. These instruments can then be adapted with various other instruments to fractionate the sample (e.g., liquid chromatography or solid-phase adsorption techniques based on chemical or biological features) and ionize the sample for introduction into the mass spectrometer (including matrix-assisted laser desorption (MALDI), electrospray, or nanospray ionization (ESI), or combinations thereof).

[0093] Generally, any mass spectrometry (MS) technique (e.g., tandem mass spectrometry, MS / MS; or post-source decay, TOF MS) that can provide accurate information about peptide mass, preferably also about the fragmentation and / or (partial) amino acid sequence of selected peptides, can be used in the methods disclosed herein. Suitable peptide MS and MS / MS techniques and systems are well known per se (see, for example, Methods in Molecular Biology, Vol. 146: "Mass Spectrometry of Proteins and Peptides", Chapman, ed., Humana Press, 2000; Biemann, 1990, Methods Enzymol, Vol. 193: pp. 455-79; or Methods in Enzymology, Vol. 402: "Biological Mass Spectrometry", Burlingame, ed., Academic Press, 2005) and can be used in carrying out the methods disclosed herein. Thus, in some embodiments, the disclosed methods comprise performing quantitative MS to measure one or more biomarkers. Such quantitative methods can be performed in an automated (Villanueva et al., Nature Protocols (2006) 1(2):880-891) or semi-automated format. In certain embodiments, the MS can be operably linked to a liquid chromatography device (LC-MS / MS or LC-MS) or a gas chromatography device (GC-MS or GC-MS / MS). Other methods useful in this context include isotope-coded affinity tagging (ICAT), tandem mass tagging (TMT), or stable isotope labeling with amino acids in cell culture (SILAC), followed by chromatography and MS / MS.

[0094] As used herein, the terms "multiple reaction monitoring (MRM)" or "selected reaction monitoring (SRM)" refer to MS-based quantification methods that are particularly useful for quantifying low-abundance analytes. In an SRM experiment, a predefined precursor ion and one or more of its fragments are selected by two mass filters in a triple quadrupole instrument and monitored over time for accurate quantification. Multiple SRM precursor and fragment ion pairs can be measured within the same experiment on a chromatographic timescale by rapidly toggling between different precursor / fragment pairs to perform an MRM experiment. A series of transitions (precursor / fragment ion pairs) combined with the retention time of a target analyte (e.g., a peptide or small molecule, such as a chemical, steroid, hormone, etc.) can constitute the final assay. Multiple analytes can be quantified during a single LC-MS experiment. The terms "schedule" or "dynamic" in reference to MRM or SRM refer to a variation of the assay in which transitions for a particular analyte are acquired only in a time window surrounding the expected retention time, significantly increasing the number of analytes that can be detected and quantified in a single LC-MS experiment and contributing to the selectivity of the test. This is because retention time is a characteristic that depends on the physical properties of the analyte. A single analyte can also be monitored using more than one transition. Finally, a standard corresponding to the analyte of interest (e.g., the same amino acid sequence) but differing due to the inclusion of a stable isotope can be included in the assay. A stable isotope standard (SIS) can be incorporated into the assay at precise levels and used to quantify the corresponding unknown analyte. An additional level of specificity is contributed by the coelution of an unknown analyte and its corresponding SIS and the characteristics of their transitions (e.g., similarity in the ratio of the levels of two transitions in the unknown and the ratio of two transitions in the corresponding SIS).

[0095] Suitable mass spectrometry assays, instruments, and systems for biomarker peptide analysis include, but are not limited to, matrix-assisted laser desorption / ionization time-of-flight (MALDI-TOF) MS; MALDI-TOF post-source decay (PSD); MALDI-TOF / TOF; surface-enhanced laser desorption / ionization time-of-flight mass spectrometry (SELDI-TOF) MS; electrospray ionization mass spectrometry (ESI-MS); ESI-MS / MS; ESI-MS / (MS) n (n is an integer greater than zero); ESI 3D or linear (2D) ion trap MS; ESI triple quadrupole MS; ESI quadrupole orthogonal time-of-flight (Q-TOF); ESI Fourier transform MS system; desorption / ionization on silicon (DIOS); secondary ion mass spectrometry (SIMS); atmospheric pressure chemical ionization mass spectrometry (APCI-MS); APCI-MS / MS; APCI-(MS) n ;Ion mobility spectroscopy (IMS);Inductively coupled plasma mass spectrometry (ICP-MS);Atmospheric pressure photoionization mass spectrometry (APPI-MS);APPI-MS / MS, and APPI-(MS) nFragmentation of peptide ions in tandem MS (MS / MS) configurations can be achieved using techniques established in the art, such as collision-induced dissociation (CID). As described herein, detection and quantification of biomarkers by mass spectrometry can include, for example, multiple reaction monitoring (MRM), as described by Kuhn et al., Proteomics 4:1175-86 (2004), among others. Scheduled multiple reaction monitoring (scheduled MRM) mode acquisition during LC-MS / MS analysis enhances the sensitivity and accuracy of peptide quantification. Anderson and Hunter, Molecular and Cellular Proteomics 5(4):573 (2006). As described herein, mass spectrometry-based assays can be advantageously combined with upstream peptide or protein separation or fractionation methods, such as chromatography and other methods described herein below. As further described herein, shotgun quantitative proteomics can be combined with SRM / MRM-based assays for high-throughput identification and validation of prognostic biomarkers for preterm birth. Antibodies that bind to SHBG

[0096] The present specification describes an isolated anti-SHBG antibody or its antigen-binding fragment, a polypeptide and its fragment, a nucleic acid and expression vector encoding the antibody, a recombinant cell containing the vector, and a composition comprising the antibody.The antibody disclosed herein has one or more desirable functional properties, including but not limited to high affinity binding to SHBG and specific binding to SHBG, which provides the ability to detect, diagnose, or treat certain conditions as described herein.The antibody disclosed herein is also useful for assessing health status or predicting adverse pregnancy events.

[0097] In one aspect, the present disclosure relates to an isolated anti-SHBG antibody or antigen-binding fragment thereof. In some embodiments, the isolated anti-SHBG antibody or fragment thereof binds to an SHBG epitope. The SHBG epitope may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or 100% identity to SEQ ID NO: 110. In some embodiments, the isolated anti-SHBG antibody or fragment thereof binds to an SHBG epitope. The SHBG epitope may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or 100% identity to SEQ ID NO: 120. In some embodiments, the isolated anti-SHBG antibody or antigen-binding antibody comprises a heavy chain, wherein the heavy chain comprises heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 11-22, 115, and 116. In some embodiments, the isolated anti-SHBG antibody or antigen-binding antibody comprises a light chain, wherein the light chain comprises light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-34. In some embodiments, the isolated anti-SHBG antibody or antigen-binding antibody comprises a heavy chain comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3, and / or light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3, comprising any one or more amino acid sequences set forth in Table 14 or Table 16. In some embodiments, the anti-SHBG antibodies provided herein comprise or consist of six CDRs. For example, in some embodiments, the anti-SHBG antibodies provided herein comprise HCDR1, HCDR2, and HCDR3, wherein the HCDR comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 11-22, 44, 45, 47, 48, 50-54, 115, or 117, and further comprise a light chain, wherein the light chain comprises LCDR1, LCDR2, and LCDR3, each comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-34.In other embodiments, the anti-SHBG antibody provided herein comprises less than six CDRs of the anti-SHBG antibody described herein. In some embodiments, the heavy chain associates with the light chain to form a binding site for SHBG. In some embodiments, the antibody provided herein comprises a heavy chain variable region. In other embodiments, the antibody provided herein comprises a light chain variable region or a VL chain. In some embodiments, the antibody provided herein has a combination of (i) a heavy chain variable region; and / or (ii) a light chain variable region.

[0098] In some embodiments, the antibodies provided herein comprise or consist of six CDRs, e.g., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and / or LCDR3, as identified in Tables 14 and 16. In some embodiments, the antibodies provided herein may comprise fewer than six CDRs. In some embodiments, the antibodies comprise or consist of one, two, three, four, or five CDRs selected from the group consisting of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and / or LCDR3, as identified in Tables 14 and 16. In some embodiments, the antibody comprises or consists of one, two, three, four, or five CDRs selected from the group consisting of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and / or LCDR3 of a monoclonal antibody selected from the group consisting of: (a) the antibody designated 13C9.E5; (b) the antibody designated variant 13C9.E5; (c) the antibody designated 4G10.F8; or (d) the antibody designated variant 4G10.F8. Thus, in some embodiments, the antibody comprises or consists of one, two, three, four, or five CDRs of any one of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and / or LCDR3 identified in Tables 14 and 16.

[0099] In some embodiments, the antibodies provided herein comprise one or more (e.g., one, two, or three) HCDRs listed in Tables 14 and 16. In other embodiments, the antibodies provided herein comprise one or more (e.g., one, two, or three) LCDRs listed in Tables 14 and 16. In yet other embodiments, the antibodies provided herein comprise one or more (e.g., one, two, or three) HCDRs listed in Tables 14 and 16, and one or more LCDRs listed in Tables 14 and 16.

[0100] Thus, in certain embodiments, the antibody comprises a variable heavy (VH) chain CDR1 having the amino acid sequence of any one of SEQ ID NOs: 11, 14, 17, 20, 52, and 115. In another embodiment, the antibody comprises a VH CDR2 having the amino acid sequence of any one of SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117. In another embodiment, the antibody comprises a VH CDR3 having the amino acid sequence of any one of SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54. In certain embodiments, the antibody comprises a VH CDR1 and / or VH CDR2 and / or VH CDR3 independently selected from the VH CDR1, VH CDR2, VH CDR3 represented in any one of the amino acid sequences represented in Table 14 and Table 16. In certain embodiments, the antibody comprises a VL CDR1 having the amino acid sequence of any one of SEQ ID NOs: 23, 26, 29, and 32. In another embodiment, the antibody comprises a variable light (VL) chain CDR2 having the amino acid sequence of any one of SEQ ID NOs: 24, 27, 30, and 33. In another embodiment, the antibody comprises a VL CDR3 having the amino acid sequence of any one of SEQ ID NOs: 25, 31, and 34. In certain embodiments, the antibody comprises a VL CDR1 and / or a VL CDR2 and / or a VL CDR3 independently selected from the VL CDR1, VL CDR2, VL CDR3 depicted in any one of the amino acid sequences depicted in Table 14 and Table 16.

[0101] Also provided herein are antibodies comprising one or more (e.g., one, two, or three) VH CDRs and one or more (e.g., one, two, or three) VL CDRs listed in Table 14 and Table 16. In particular, VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115) and VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32) listed in Table 14 and Table 16; VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115) and VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115) and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117) and VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32); VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117) and VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33); VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117) and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54) and VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32); VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54) and VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33); VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54) and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117) and VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117) and VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33);VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54), and VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54) and VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33); VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54) and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32) and VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32) and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33) and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32) and VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33); VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32) and VL CDR3 (SEQ ID NOs: 25, 31, and 34);VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33), and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32), and VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33); VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32), and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54) and VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54), and VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54), and VL CDR3 (SEQ ID NOs: 25, 31, and 34);VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32), and VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32), and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33), and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32), and VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH VL CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54), VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33), and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54) ), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32) and VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33);VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32), and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54), VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33), and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32), and VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54) ), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32) and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54), VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33) and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32), VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33), and VL CDR3 (SEQ ID NOs: 25, 31, and 34);VH CDR1 (SEQ ID NOs: 11, 14, 17, 20, 52, and 115), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32), VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33), and VL CDR3 (SEQ ID NOs: 25, 31, and 34); VH CDR2 (SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117), VH CDR3 (SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54), VL CDR1 (SEQ ID NOs: 23, 26, 29, and 32), VL CDR2 (SEQ ID NOs: 24, 27, 30, and 33), and VL CDR3 (SEQ ID NOs: 25, 31, and 34); or VH CDR (SEQ ID NOs: Provided herein are antibodies comprising any combination of VL CDRs (SEQ ID NOs: 23, 24, 25, 26, 27, 29, 30, 31, 32, 33) and VL CDRs (SEQ ID NOs: 24, 25, 26, 27, 29, 30, 31, 32, 33).

[0102] In some embodiments, the isolated anti-SHBG antibody or antigen binding comprises a heavy chain, wherein the heavy chain comprises a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 11-22, 44, 45, 47, 48, 50-54, 115, or 117. In some embodiments, the isolated anti-SHBG antibody or antigen binding comprises a light chain comprising a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-34. In some embodiments, the heavy chain associates with the light chain to form a binding site for SHBG.

[0103] The anti-SHBG antibody or antigen-binding fragment thereof can comprise a heavy chain comprising an amino acid sequence having at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 36, 124, or 126. In some embodiments, the anti-SHBG antibody or antigen-binding fragment thereof can comprise a heavy chain variable region comprising an amino acid sequence having at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 8 and 40. In some embodiments, the anti-SHBG antibody or antigen-binding fragment thereof can comprise a light chain comprising an amino acid sequence having at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 38, 125, or 127. In some embodiments, the anti-SHBG antibody or antigen-binding fragment thereof can comprise a light chain variable region comprising an amino acid sequence having at least 85%, preferably 90%, more preferably 95% or more, e.g., 95%, 96%, 97%, 98%, or 99% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 and 42. In some embodiments, the heavy chain associates with the light chain to form a binding site for SHBG. In some embodiments, the anti-SHBG antibody or antigen-binding fragment thereof is humanized.

[0104] In some embodiments, the anti-SHBG antibody or antigen-binding fragment is monoclonal. In some embodiments, the anti-SHBG antibody or antigen-binding fragment is a FAB, Fab', F(ab')2, Fv, scFv, (scFv)2, single-chain antibody molecule, dual variable region antibody, single variable region antibody, linear antibody, or V region formed from an antibody fragment. In some embodiments, the anti-SHBG antibody or antigen-binding fragment is humanized, human, or chimeric.

[0105] In some embodiments, the anti-SHBG antibody is conjugated or recombinantly fused to a diagnostic or detectable agent. In some embodiments, the conjugated or recombinantly bound antibody comprises a masked or activatable conjugate and may be useful, for example, for preventing a disease or disorder, such as preterm birth, preeclampsia, or any other pregnancy-related disorder. The conjugated or recombinantly fused anti-SHBG antibody or antigen-binding fragment thereof may be useful, for example, for monitoring or prognosing the onset, development, progression, and / or severity of preterm birth, preeclampsia, or any other pregnancy-related disorder.

[0106] In some embodiments, diagnostic or detectable agents include, for example, enzymes, including but not limited to, horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase; prosthetic groups, including but not limited to, streptavidin / biotin or avidin / biotin; fluorescent materials, including but not limited to, umbelliferone, fluorescein, fluorescein isothiocynate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride, or phycoerythrin; luminescent materials, including but not limited to, luminol; bioluminescent materials, including but not limited to, luciferase, luciferin, or aequorin; chemiluminescent materials, including but not limited to, acridinium-based compounds or HALOTAG; These include, but are not limited to, iodine (131I, 125I, 123I, and 121I), carbon (14C), sulfur (35S), tritium (3H), indium (115In, 113In, 112In, and 111In), technetium (99Tc), thallium (201Ti), gallium (68Ga and 67Ga), palladium (103Pd), molybdenum (99Mo), xenon (133Xe), fluorine (18F), 153Sm, 177 Radioactive materials include Lu, Gd, Pm, La, Yb, Ho, Y, Sc, Re, Re, Pr, Rh, Ge, Co, Zn, Sr, P, Gd, Yb, Cr, Mn, Se, Sn, or Sn; positron-emitting metals using various positron emission tomography techniques; and non-radioactive paramagnetic metal ions.

[0107] In another aspect, the present disclosure relates to binding agents that bind to the same epitope as the anti-SHBG antibodies or antigen-binding fragments disclosed herein. In some embodiments, the SHBG binding agent comprises a polypeptide or fragment thereof that is not an antibody.

[0108] In another aspect, the present disclosure relates to an isolated nucleic acid encoding the anti-SHBG antibody or its antigen-binding fragment disclosed herein.Those skilled in the art will recognize that the coding sequence of a protein can be changed (e.g., replaced, deleted, inserted, etc.) without changing the amino acid sequence of the protein.Therefore, those skilled in the art will understand that the nucleic acid sequence encoding the anti-SHBG antibody disclosed herein can be modified without changing the amino acid sequence of the protein.

[0109] In another aspect, the present disclosure relates to nucleic acid molecules encoding the heavy and / or light chains of the antibodies or fragments thereof disclosed herein. The nucleic acid molecules are preferably isolated nucleic acid molecules. Isolated nucleic acid molecules can be substantially pure nucleic acid molecules. In some embodiments, the nucleic acid molecules encode the heavy chain of an anti-SHBG antibody or fragment thereof. In some embodiments, the nucleic acid molecule encoding the heavy chain has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to at least one of SEQ ID NOs: 3, 35, 200, or 202. In some embodiments, the nucleic acid molecules encode the light chain of an anti-SHBG antibody or fragment thereof. In some embodiments, the nucleic acid molecule encoding the light chain has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NOs: 5, 37, 201, or 203.

[0110] In another aspect, the present disclosure relates to a vector comprising the isolated nucleic acid disclosed herein. Any vector known to one of skill in the art in light of the present disclosure can be used, for example, a plasmid, cosmid, phage vector, or viral vector. In some embodiments, the vector is a recombinant expression vector, for example, a plasmid. The vector can include any elements for establishing the conventional functions of an expression vector, such as a promoter, a ribosome-binding element, a terminator, an enhancer, a selection marker, and an origin of replication. The promoter can be a constitutive promoter, an inducible promoter, or a repressible promoter. Several expression vectors capable of delivering nucleic acids to cells are known in the art and can be used herein for producing antibodies or antigen-binding fragments thereof in cells. Conventional cloning techniques or artificial gene synthesis can be used to generate recombinant expression vectors according to the embodiments disclosed herein. Such techniques are well known to one of skill in the art in light of the present disclosure.

[0111] In another aspect, the present disclosure relates to a host cell comprising an isolated nucleic acid encoding the anti-SHBG antibody or antigen-binding fragment thereof disclosed herein. Any host cell known to those skilled in the art in light of the present disclosure can be used for recombinant expression of the antibodies or antigen-binding fragments thereof disclosed herein. In some embodiments, the host cell is E. coli TG1 or BL21 cell (e.g., for expression of scFv or Fab antibodies), CHO-DG44 or CHO-K1 cell, or HEK293 cell (e.g., for expression of full-length IgG antibodies). According to certain embodiments, the recombinant expression vector is transformed into the host cell by conventional methods, such as chemical transfection, heat shock, or electroporation, which stably integrates the recombinant nucleic acid into the genome of the host cell so that it is efficiently expressed.

[0112] In another aspect, the present disclosure relates to a method for producing an anti-SHBG antibody or antigen-binding fragment thereof disclosed herein. The method includes culturing cells containing a nucleic acid encoding the anti-SHBG antibody or antigen-binding fragment thereof disclosed herein under conditions to produce the anti-SHBG antibody or antigen-binding fragment thereof, and recovering the antibody or antigen-binding fragment thereof from the cells or cell culture (e.g., from the supernatant). The expressed antibody or antigen-binding fragment thereof can be recovered and purified from the cells according to conventional techniques known in the art and described herein.

[0113] In another aspect, the present disclosure relates to a buffer composition comprising an isolated anti-SHBG antibody or antigen-binding fragment thereof provided herein. In some embodiments, the buffer composition may comprise at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or more by weight of the anti-SHBG antibody or other binding agent having an antigen-binding fragment that specifically binds to SHBG. In other embodiments, for example, the anti-SHBG antibody or other binding agent having an antigen-binding fragment that specifically binds to SHBG may comprise about 2% to about 75%, about 25% to about 60%, about 50% to about 50%, or any range therein by weight of the composition.

[0114] In some embodiments, the anti-SHBG antibodies provided herein bind to SHBG (e.g., human SHBG) at a binding affinity of ≦1 μM, ≦0.5 μM, ≦0.1 μM, ≦0.01 μM, ≦0.001 μM, ≦100 nM, ≦10 nM, ≦2 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM (e.g., 10 -8 M or less, e.g., 10 -8 M~10 -13 M, e.g., 10 -9 M~10 -13 Dissociation constant (K DIn some embodiments, the anti-SHBG antibodies provided herein bind to SHBG (e.g., human SHBG) at a binding affinity of about 1 x 10 -7 M or less, 1 x 10 -8 M or less, 1 x 10 -9 M or less, 1 x 10 -10 M or less, 2 x 10 -7 M or less, 2 x 10 -8 M or less, 2 x 10 -9 M or less, 2 x 10 -10 M or less, 5 x 10 -7 M or less, 5 x 10 -8 M or less, 5 x 10 -9 M or less, or 5 x 10 -10 M or less K D For example, the anti-SHBG antibodies described herein bind at approximately 1.0 x 10 -9 M ~ approx. 2.5×10 -9 K of M D In some embodiments, the anti-SHBG antibody may have a K of about 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, or 3.0 nM. D In some embodiments, the anti-SHBG antibody has a K of about 5.0, 6.0, 7.0, 8.0, 9.0, or 10 nM. D In some embodiments, the anti-SHBG antibody has a K of about 15, 20, or 25 nM. D In certain embodiments, the anti-SHBG antibody has a K of about 22 nM. DA variety of methods for measuring binding affinity are known in the art, any of which can be used for purposes of the present disclosure, including, for example, by RIA performed using the antibody of interest and a Fab version of its antigen (Chen et al., 1999, J. Mol Biol 293:865-81); by Octet®, e.g., using the Octet® Red 96 system, or by Biacore®, e.g., using a Biacore® TM-2000 or Biacore® TM-3000, biolayer interferometry (BLI) or surface plasmon resonance (SPR) assays. "On-rate" or "rate of association" or "association rate" or "k" can also be determined using the same biolayer interferometry (BLI) or surface plasmon resonance (SPR) techniques described above, for example, using Octet® Red96, Biacore® TM-2000, or Biacore® TM-3000 systems.

[0115] In some embodiments, the anti-SHBG antibodies provided herein are those described in Examples 1 and 2 below. Thus, in some embodiments, the antibodies provided herein comprise one or more CDR sequences of any one of SEQ ID NOS: 11-34, 44, 45, 47, 48, 50-54, 115, or 117. CDR sequences can be determined according to well-known numbering systems. In some embodiments, the CDRs follow the ImMunoGeneTics (IMGT) numbering system. IMGT is an integrated information system specialized in immunoglobulins (IGs), T cell receptors (TRs), and major histocompatibility complexes (MHCs) of humans and other vertebrates. Herein, CDRs are referred to in terms of both amino acid sequence and location within the light or heavy chain. Because the "location" of CDRs within the structure of immunoglobulin variable domains is conserved across species and occurs in structures called loops, CDR and framework residues are readily identified by using a numbering system that aligns variable domain sequences according to structural features. Thus, in some embodiments, the isolated anti-SHBG antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 11, 12, and 13, respectively. In some embodiments, the isolated anti-SHBG antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 11, 44, and 45, respectively. In some embodiments, the isolated anti-SHBG antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 23, 24, and 25, respectively. In some embodiments, the HCDR associates with the LCDR to form an SHBG binding region.

[0116] In some embodiments, the CDRs are determined according to Kabat numbering (see, e.g., Kabat et al., 5 th(See Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). The Kabat numbering scheme is a scheme for numbering amino acid residues in antibodies based on the variable region. The Kabat numbering of residues can be determined for a given antibody by alignment of the antibody's sequence with a "standard Kabat numbering sequence" in regions of homology. The Kabat numbering system is generally used when referring to residues in the variable domain. Thus, in some embodiments, an isolated anti-SHBG antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 115, 116, and 19, respectively. In some embodiments, an isolated anti-SHBG antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 115, 117, and 51, respectively. In some embodiments, an isolated anti-SHBG antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 29, 30, and 31, respectively. In some embodiments, the HCDR associates with the LCDR to form an SHBG binding region.

[0117] In some embodiments, CDRs are determined according to Chothia numbering. Chothia refers to the location of structural loops (see, e.g., Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). Thus, in some embodiments, an isolated anti-SHBG antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 14, 15, and 16, respectively. In some embodiments, an isolated anti-SHBG antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 14, 47, and 48, respectively. In some embodiments, an isolated anti-SHBG antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 26, 27, and 28, respectively. In some embodiments, an isolated anti-SHBG antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 26, 27, and 25, respectively. In some embodiments, the HCDR associates with the LCDR to form an SHBG binding region.

[0118] In some embodiments, the CDRs are determined according to AbM numbering. AbM hypervariable regions represent a compromise between Kabat CDRs and Chothia structural loops and are used by Oxford Moleculars AbM antibody modeling software (see, e.g., Martin, in Antibody Engineering, Vol. 2, Chapter 3, Springer Verlag). Thus, in some embodiments, the isolated anti-SHBG antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 17, 18, and 19, respectively. In some embodiments, the isolated anti-SHBG antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 17, 50, and 51, respectively. In some embodiments, the isolated anti-SHBG antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 29, 30, and 31, respectively. In some embodiments, the HCDR associates with the LCDR to form an SHBG binding region.

[0119] In some embodiments, the CDRs are determined according to Contact numbering. Thus, in some embodiments, the isolated anti-SHBG antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 20, 21, and 22, respectively. In some embodiments, the isolated anti-SHBG antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 52, 53, and 54, respectively. In some embodiments, the isolated anti-SHBG antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 32, 33, and 34, respectively. In some embodiments, the HCDR associates with the LCDR to form an SHBG binding region.

[0120] In other embodiments, the CDRs follow Contact numbering. In some embodiments, the anti-SHBG antibody is humanized. In some embodiments, the anti-SHBG antibody is partially humanized. In some embodiments, the anti-SHBG antibody is chimeric. In some embodiments, the anti-SHBG antibody is a humanized chimeric. In some embodiments, the anti-SHBG antibody is a murine antibody. In some embodiments, the anti-SHBG antibody comprises an acceptor human framework, e.g., a human immunoglobulin framework or a human consensus framework. In some embodiments, the anti-SHBG antibody comprises an acceptor murine framework, e.g., a murine immunoglobulin framework or a murine consensus framework.

[0121] In some embodiments, the anti-SHBG antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 11-13. In some embodiments, the anti-SHBG antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 14-16. In some embodiments, the anti-SHBG antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 17-19. In some embodiments, the anti-SHBG antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 20-22. In some embodiments, the anti-SHBG antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 115, 116, and 19. In some embodiments, the anti-SHBG antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 11, 44, and 45. In some embodiments, the anti-SHBG antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 14, 47, and 48. In some embodiments, the anti-SHBG antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 17, 50, and 51. In some embodiments, the anti-SHBG antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 52, 53, and 54. In some embodiments, the anti-SHBG antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 115, 117, and 51. The CDR sequences can be determined according to well-known numbering systems or combinations thereof. In some embodiments, the CDRs are according to IMGT numbering. In some embodiments, the CDRs are according to Kabat numbering. In some embodiments, the CDRs are according to AbM numbering. In other embodiments, the CDRs are according to Chothia numbering. In other embodiments, the CDRs are according to Contact numbering.

[0122] In some embodiments, the anti-SHBG antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 23, 24, and 25. In some embodiments, the anti-SHBG antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 26, 27, and 28. In some embodiments, the anti-SHBG antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 29, 30, and 31. In some embodiments, the anti-SHBG antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 32, 33, and 34. In some embodiments, the anti-SHBG antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 29, 30, and 31. In some embodiments, the anti-SHBG antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 26, 27, and 25. CDR sequences can be determined according to known numbering systems or their combinations.In some embodiments, CDRs are according to IMGT numbering.In some embodiments, CDRs are according to AbM numbering.In other embodiments, CDRs are according to Chothia numbering.In other embodiments, CDRs are according to Contact numbering.

[0123] In some embodiments, the antibodies or fragments thereof provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 11, 12, and 13, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 23, 24, and 25. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 14, 15, and 16, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 26, 27, and 28. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 17, 18, and 19, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 29, 30, and 31. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 20, 21, and 22, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 32, 33, and 34. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 115, 116, and 19, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 29, 30, and 31. Thus, the CDR sequences can be determined according to well-known numbering systems or combinations thereof. In some embodiments, the CDRs are according to IMGT numbering. In some embodiments, the CDRs are according to AbM numbering. In other embodiments, the CDRs are according to Chothia numbering. In other embodiments, the CDRs are according to Contact numbering.

[0124] In some embodiments, the antibodies or fragments thereof provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 11, 44, and 45, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 23, 24, and 25. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 14, 47, and 48, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 26, 27, and 25. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 17, 50, and 51, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 29, 30, and 31. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 52, 53, and 54, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 32, 33, and 34. In some embodiments, the antibody or antigen-binding fragment provided herein comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 115, 117, and 51, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 29, 30, and 31. Thus, the CDR sequences can be determined according to well-known numbering systems or combinations thereof. In some embodiments, the CDRs are according to IMGT numbering. In some embodiments, the CDRs are according to AbM numbering. In other embodiments, the CDRs are according to Chothia numbering. In other embodiments, the CDRs are according to Contact numbering.

[0125] In some embodiments, an HCDR1 comprises an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 11, 14, 17, 20, and 115; (ii) an HCDR1 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 12, 15, 18, 21, and 116; (iii) an HCDR3 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 13, 16, 19, and 22; (iv) SEQ ID NOs: 23, 26, (v) an LCDR1 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 24, 27, 30, and 33; Provided herein is an antibody or fragment thereof that binds to SHBG, comprising: (vi) an LCDR2 comprising an amino acid sequence with 8%, 99%, or 100% sequence identity; and / or (vii) an LCDR3 comprising an amino acid sequence with at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of SEQ ID NOs: 25, 28, 31, and 34. In some embodiments, the anti-SHBG antibody is humanized. In some embodiments, the anti-SHBG antibody comprises an acceptor human framework, such as a human immunoglobulin framework or a human consensus framework.

[0126] In some embodiments, (ii) an HCDR1 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 11, 14, 17, 52, and 115; , 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 45, 48, 51, and 54; (iii) HCDR3 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 45, 48, 51, and 54; (iv) SEQ ID NOs: 23, 26, 29 , and 32; (v) an LCDR1 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 24, 27, 30, and 33; Provided herein is an antibody or fragment thereof that binds to SHBG, comprising: (vi) an LCDR2 comprising an amino acid sequence with 8%, 99%, or 100% sequence identity; and / or (vii) an LCDR3 comprising an amino acid sequence with at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of SEQ ID NOs: 25, 31, and 34. In some embodiments, the anti-SHBG antibody is humanized. In some embodiments, the anti-SHBG antibody comprises an acceptor human framework, such as a human immunoglobulin framework or a human consensus framework.

[0127] In one aspect, the anti-SHBG antibodies or fragments thereof disclosed herein may contain some variation in HCDR2 and / or HCDR3. Thus, provided herein is an isolated anti-SHBG antibody or fragment thereof comprising a heavy chain comprising an HCDR1 comprising the amino acid sequence GYTFTGYW (SEQ ID NO: 11) and an HCDR2 comprising the amino acid sequence IDPSX1X2YT (SEQ ID NO: 112). The residue identified as X1 can independently be any naturally occurring amino acid. Similarly, the residue identified as X2 can independently be any naturally occurring amino acid residue. The anti-SHBG antibody or fragment thereof can further comprise an HCDR3 comprising the amino acid sequence TRX3PLVTADFX3Y (SEQ ID NO: 113). The residue identified as X3 can independently be any naturally occurring amino acid residue. In some embodiments, X1 comprises threonine or aspartic acid. In some embodiments, X2 comprises glycine or threonine. In some embodiments, X3 comprises glutamic acid or aspartic acid. In some embodiments, the antibody or fragment thereof, X1 is threonine, X2 is glycine, and X3 is glutamic acid. In some embodiments, the anti-SHBG antibody or fragment thereof comprises a light chain comprising an LCDR1 comprising the amino acid sequence QNVDTN (SEQ ID NO: 23), an LCDR2 comprising the amino acid sequence SAS (SEQ ID NO: 24), and an LCDR3 comprising the amino acid sequence QQYDSYPY (SEQ ID NO: 25).

[0128] In some embodiments, the anti-SHBG antibody or fragment thereof has a higher affinity for a biologically active form of the SHBG protein than for a biologically inactive form of SHBG. In some embodiments, biologically active SHBG is a version of SHBG that can enter cells or bind to a target receptor. In some embodiments, biologically active SHBG includes a version of SHBG that is glycosylated. Thus, in some embodiments, the anti-SHBG antibody or fragment thereof disclosed herein includes a higher affinity for glycosylated SHBG than for non-glycosylated SHBG. In some embodiments, the anti-SHBG antibody or fragment thereof can differentially bind to an SHBG protein that includes an epitope that includes a glycosylated amino acid, where the glycosylated amino acid includes a threonine at position 36. In some embodiments, the glycosylated amino acid includes an O-linked glycan.

[0129] In some embodiments, the antibody provided herein is humanized antibody.The framework region described herein is determined based on the boundary of CDR numbering system.In other words, when CDR is determined by Kabat, IMGT or Chothia for example, framework region is the amino acid residue around the CDR of variable region from N-terminus to C-terminus in the format: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. For example, FR1 is defined as the amino acid residues N-terminal to the CDR1 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; FR2 is defined as the amino acid residues between the CDR1 and CDR2 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; FR3 is defined as the amino acid residues between the CDR2 and CDR3 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; and FR4 is defined as the amino acid residues C-terminal to the CDR3 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system.

[0130] In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a VH sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to the sequence of SEQ ID NO:8, and a VL sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO:10. In some embodiments, an antibody or antigen-binding fragment provided herein comprises a VH sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 40, and a VL sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 42.

[0131] In certain embodiments, the antibodies or antigen-binding fragments thereof described herein comprise an amino acid sequence having a certain percent identity to any of the antibodies provided herein, for example, those set forth in the sequence listing below.

[0132] The percent identity between two sequences (for example, amino acid sequence or nucleic acid sequence) can be determined by mathematical algorithm.A non-limiting example of the mathematical algorithm used for comparing two sequences is the algorithm of Karlin and Altschul, Proc. Natl. Acad. Sci. USA 87:2264 2268 (1990), modified as in Karlin and Altschul, Proc. Natl. Acad. Sci. USA 90:5873 5877 (1993).Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul et al., J. Mol. Biol. 215:403 (1990).BLAST nucleotide search can be performed by setting the parameters of NBLAST nucleotide program, for example, score=100, word length=12, to obtain the nucleotide sequence homologous to the nucleic acid molecules described herein. BLAST protein searches can be performed by setting the parameters of the XBLAST program, for example, to score 50 and word length = 3, to obtain amino acid sequences homologous to the protein molecules described herein. To obtain gapped alignments for comparison purposes, Gapped BLAST can be used, as described in Altschul et al., Nucleic Acids Res. 25:3389 3402 (1997). Alternatively, PSI BLAST can be used to perform an iterated search that detects distant relationships between molecules (ibid.). When using BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of each program (e.g., XBLAST and NBLAST) can be used (see, for example, the National Center for Biotechnology Information (NCBI) on the World Wide Web at ncbi.nlm.nih.gov).Another non-limiting example of a mathematical algorithm utilized for comparing sequences is the algorithm of Myers and Miller, CABIOS 4:11-17 (1998). Such an algorithm is incorporated into the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package. When utilizing the ALIGN program to compare amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used. The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percent identity, typically, only perfect matches are counted.

[0133] In some embodiments, the antibodies provided herein contain substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but the anti-SHBG antibody comprising that sequence retains the ability to bind to SHBG. In some embodiments, a total of 1 to 10 amino acids are substituted, inserted, and / or deleted in the reference amino acid sequence. In some embodiments, the substitutions, insertions, or deletions occur in regions outside the CDRs (i.e., in the FRs). Optionally, the anti-SHBG antibodies provided herein include post-translational modifications of the reference sequence.

[0134] In some embodiments, functional epitopes can be mapped, for example, by combinatorial alanine scanning, to identify amino acids in the SHBG protein that are required for interaction with the anti-SHBG antibodies provided herein. In some embodiments, the conformation and crystal structure of an anti-SHBG antibody bound to SHBG can be used to identify the epitope. In some embodiments, the present disclosure provides antibodies that specifically bind to the same epitope as any of the anti-SHBG antibodies provided herein. For example, in some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-SHBG antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 8 and a VL comprising the amino acid sequence of SEQ ID NO: 10. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-SHBG antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 40 and a VL comprising the amino acid sequence of SEQ ID NO: 42.

[0135] In some embodiments, the present invention provides an anti-SHBG antibody or its antigen-binding fragment, which specifically binds to SHBG competitively with any one of the anti-SHBG antibodies described herein.For example, in some embodiments, the present invention provides an antibody or antigen-binding fragment that binds to the same epitope as the anti-SHBG antibody that comprises VH comprising the amino acid sequence of SEQ ID NO: 8 and VL comprising the amino acid sequence of SEQ ID NO: 10.In some embodiments, the present invention provides an antibody or antigen-binding fragment that binds to the same epitope as the anti-SHBG antibody that comprises VH comprising the amino acid sequence of SEQ ID NO: 40 and VL comprising the amino acid sequence of SEQ ID NO: 42.

[0136] In some embodiments, provided herein is an SHBG binding agent comprising any one of the anti-SHBG antibodies described above. In some embodiments, the SHBG binding agent is a monoclonal antibody, including a mouse antibody, a chimeric antibody, a humanized antibody, or a human antibody. In some embodiments, the anti-SHBG antibody is an antibody fragment, such as an scFv. In some embodiments, the SHBG binding agent is a fusion protein comprising the anti-SHBG antibody provided herein. In other embodiments, the SHBG binding agent is a multispecific antibody comprising the anti-SHBG antibody provided herein. Other exemplary SHBG binding agents are described in more detail in the following sections.

[0137] In some embodiments, an anti-SHBG antibody or antigen binding protein according to any of the above embodiments may incorporate any single or combination of features, as described below. Antibodies that bind to IBP4

[0138] Described herein are isolated anti-IBP4 antibodies or antigen-binding fragments thereof, polypeptides and fragments thereof, nucleic acids and expression vectors encoding the antibodies, recombinant cells containing the vectors, and compositions containing the antibodies.The antibodies disclosed herein have one or more desirable functional properties, including, but not limited to, high affinity binding to IBP4 and specific binding to IBP4, which provides the ability to detect, diagnose, or treat certain conditions as described herein.The antibodies disclosed herein are also useful for assessing health status or predicting adverse pregnancy events.

[0139] In some embodiments, the anti-IBP4 antibodies provided herein bind to IBP4 (e.g., human IBP4) at a concentration of ≦1 μM, ≦0.5 μM, ≦0.1 μM, ≦0.01 μM, ≦0.001 μM, ≦100 nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM (e.g., 10 -8 M or less, e.g., 10 -8 M~10 -13M, e.g., 10 -9 M~10 -13 Dissociation constant (K D In some embodiments, the anti-IBP4 antibodies provided herein bind to IBP4 (e.g., human IBP4) at a concentration of about 1 x 10 -7 M or less, 1 x 10 -8 M or less, 1 x 10 -9 M or less, 1 x 10 -10 M or less, 2 x 10 -7 M or less, 2 x 10 -8 M or less, 2 x 10 -9 M or less, 2 x 10 -10 M or less, 5 x 10 -7 M or less, 5 x 10 -8 M or less, 5 x 10 -9 M or less, or 5 x 10 -10 M or less K D For example, the anti-IBP4 antibodies described herein bind at approximately 3×10 -8M ~Approx. 8×10 -8 K of M D In some embodiments, the anti-IBP4 antibody may have a K of about 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, or 3.0 nM. D In some embodiments, the anti-IBP4 antibody has a K of about 5.0, 6.0, 7.0, 8.0, 9.0, or 10 nM. D In some embodiments, the anti-IBP4 antibody has a K of about 15, 20, or 25 nM. D In certain embodiments, the anti-IBP4 antibody has a K of about 22 nM. DA variety of methods for measuring binding affinity are known in the art, any of which can be used for purposes of the present disclosure, including, for example, by RIA performed using the antibody of interest and a Fab version of its antigen (Chen et al., 1999, J. Mol Biol 293:865-81); by Octet®, e.g., using the Octet® Red 96 system, or by Biacore®, e.g., using a Biacore® TM-2000 or Biacore® TM-3000, biolayer interferometry (BLI) or surface plasmon resonance (SPR) assays. "On-rate" or "rate of association" or "association rate" or "k" can also be determined using the same biolayer interferometry (BLI) or surface plasmon resonance (SPR) techniques described above, for example, using an Octet® Red96, Biacore® TM-2000, or Biacore® TM-3000 system.

[0140] In some embodiments, the anti-IBP4 antibodies provided herein are those described in Examples 1, 3, and 4 below. In one aspect, the present disclosure relates to an isolated anti-IBP4 antibody or antigen-binding fragment thereof. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof binds to an IBP4 epitope. The IBP4 epitope may comprise a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or 100% identity to SEQ ID NO: 111 or 131.

[0141] In some embodiments, the antibodies provided herein comprise a heavy chain variable region. In other embodiments, the antibodies provided herein comprise a light chain variable region. In some embodiments, the antibodies provided herein have a combination of (i) a heavy chain variable region; and / or (ii) a light chain variable region.

[0142] In some embodiments, the antibodies provided herein comprise or consist of six CDRs, e.g., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and / or LCDR3, identified in Tables 18, 20, 24, 26, 28, 30, and 32.

[0143] In some embodiments, the antibodies provided herein may comprise fewer than six CDRs. In some embodiments, the antibodies comprise or consist of one, two, three, four, or five CDRs selected from the group consisting of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and / or LCDR3 identified in Tables 18, 20, 24, 26, 28, 30, and 32. In some embodiments, the antibodies are selected from the group consisting of (a) the antibody designated 5B3.G9; (b) the antibody designated variant 5B3.G9; (c) the antibody designated 7D6.G4; (d) the antibody designated variant 7D6.G4; (e) the antibody designated 1H9.B6; (f) the antibody designated variant 1H9.B6; (g) the antibody designated 5C11.A7; (h) the antibody designated variant 5C11.A7; (i) the antibody designated variant 7G11.H4; (j) the antibody designated variant 7G (k) the antibody designated 3D4.E9; (l) the antibody designated variant 3D4.E9; (m) the antibody designated 1G3.A3; or (n) the antibody designated variant 1G3.A3. Thus, in some embodiments, the antibody comprises or consists of one, two, three, four, or five CDRs selected from the group consisting of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and / or LCDR3 of any one of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and / or LCDR3 identified in Tables 18, 20, 24, 26, 28, 30, and 32.

[0144] In some embodiments, the antibodies provided herein comprise one or more (e.g., one, two, or three) VH CDRs listed in Table 18, Table 20, Table 24, Table 26, Table 28, Table 30, and Table 32. In other embodiments, the antibodies provided herein comprise one or more (e.g., one, two, or three) VL CDRs listed in Table 18, Table 20, Table 24, Table 26, Table 28, Table 30, and Table 32. In yet other embodiments, the antibodies provided herein comprise one or more (e.g., one, two, or three) VH CDRs listed in Table 18, Table 20, Table 24, Table 26, Table 28, Table 30, and Table 32, and one or more VL CDRs listed in Table 18, Table 20, Table 24, Table 26, Table 28, Table 30, and Table 32. Thus, in certain embodiments, the antibody comprises a VH CDR1 having the amino acid sequence of any one of SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314. In another embodiment, the antibody comprises a VH CDR2 having the amino acid sequence of any one of SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321. In another embodiment, the antibody comprises a VH CDR3 having the amino acid sequence of any one of SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322. In certain embodiments, the antibody comprises a VH CDR1 and / or VH CDR2 and / or VH CDR3 independently selected from the VH CDR1, VH CDR2, VH CDR3 represented in any one of the amino acid sequences represented in Table 18, Table 20, Table 24, Table 26, Table 28, Table 30, and Table 32. In certain embodiments, the antibody comprises a VL CDR1 having the amino acid sequence of any one of SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330.In another embodiment, the antibody comprises a VL CDR2 having the amino acid sequence of any one of SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331. In another embodiment, the antibody comprises a VL CDR3 having the amino acid sequence of any one of SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332. In certain embodiments, the antibody comprises a VL CDR1 and / or a VL CDR2 and / or a VL CDR3 independently selected from the VL CDR1, VL CDR2, VL CDR3 represented in any one of the amino acid sequences represented in Table 18, Table 20, Table 24, Table 26, Table 28, Table 30, and Table 32.

[0145] Also provided herein are antibodies comprising one or more (e.g., one, two, or three) VH CDRs and one or more (e.g., one, two, or three) VL CDRs listed in Table 18, Table 20, Table 24, Table 26, Table 28, Table 30, and Table 32. In particular, antibodies comprising one or more (e.g., one, two, or three) VH CDRs and one or more (e.g., one, two, or three) VL CDRs listed in Table 18, Table 20, Table 24, Table 26, Table 28, Table 30, and Table 32. VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314) and VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314) and VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332); VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321) and VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330); VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321) and VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331);VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332); VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322) and VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330); VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322) and VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331); VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332);VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321) and VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321) and VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332);VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322) and VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322) and VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331); VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332);VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330) and VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332); VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330) and VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331);VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330), and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332); VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332); VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330) and VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331); VH CDR3 (SEQ ID NOs: 77, 80; 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332); VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322), VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322) and VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330);VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322) and VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332);VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330), and VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332);VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331), and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330) and VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331);VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330), and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322), VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332);VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, and 322), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330) and VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331); VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, and 322), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332);VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, and 322), VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, and 322), VL CDR1 ... VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, and 322), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332);VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322), VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331) and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332); VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330), VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331), and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332);VH CDR1 (SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330), VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331), and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332); VH CDR2 (SEQ ID NOs: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321), VH CDR3 (SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, and 322), VL CDR1 (SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330), VL CDR2 (SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331), and VL CDR3 (SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332);or VH CDRs (SEQ ID NOs: 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 118, 119, 244, 247, 250, 253, 256, 288, 291, 294, 314, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, 286, 289, 292, 295, 312, 315, 317, 319, 321, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, and 322) and VL Provided herein are antibodies comprising any combination thereof of CDRs (SEQ ID NOs: 23, 24, 89, 91, 94, 95, 97, 98, 99, 100, 101, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, 330, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, 331, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332);

[0146] In some embodiments, the antibodies provided herein comprise one or more (e.g., one, two, or three) HCDRs listed in Tables 18, 20, 24, 26, 28, 30, and 32. In other embodiments, the antibodies provided herein comprise one or more (e.g., one, two, or three) LCDRs listed in Tables 14 and 16. In still other embodiments, the antibodies provided herein comprise one or more (e.g., one, two, or three) HCDRs listed in Tables 14 and 16, and one or more LCDRs listed in Tables 18, 20, 24, 26, 28, 30, and 32.

[0147] Thus, in some embodiments, the antibodies provided herein comprise one or more CDR sequences of any one of SEQ ID NOS: 24, 75-99, 118-119, 216-240, 244-264, 272-275, 284-303, or 312-332. CDR sequences can be determined according to well-known numbering systems. In some embodiments, CDRs are determined according to ImMunoGeneTics (IMGT) numbering. IMGT is an integrated information system dedicated to immunoglobulins (IGs), T-cell receptors (TRs), and major histocompatibility complexes (MHCs) of humans and other vertebrates. Herein, CDRs are referred to in terms of both amino acid sequence and location within the light or heavy chain. Because the "location" of CDRs within the structure of immunoglobulin variable domains is conserved across species and occurs in structures called loops, CDR and framework residues are readily identified by using a numbering system that aligns variable domain sequences according to structural features. Thus, in some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 75, 76, and 77, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 75, 216, and 217, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 244, 245, and 246, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 244, 245, and 217, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 244, 284, and 285, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 244, 312, and 313, respectively.In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 87, 24, and 89, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 87, 24, and 99, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 23, 24, and 259, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 87, 24, and 259, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 226, 24, and 227, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 87, 297, and 298, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 323, 24, and 324, respectively. In some embodiments, the HCDR associates with the LCDR to form an IBP4-binding region. For example, in some embodiments, the CDR sequences can be any one or more of the CDR sequences set forth in Tables 18, 20, 24, 26, 28, 30, and 32.

[0148] In some embodiments, CDRs are determined according to Kabat numbering (see, e.g., Kabat et al., 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). The Kabat numbering scheme is a scheme for numbering amino acid residues in antibodies based on the variable region. The Kabat numbering of residues can be determined for a given antibody by alignment of the antibody's sequence with a "standard Kabat numbering sequence" in regions of homology. The Kabat numbering system is generally used when referring to residues in the variable domain. Thus, in some embodiments, an isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 118, 119, and 83, respectively. In some embodiments, an isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 118, 224, and 25, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 256, 257, and 258, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 294, 295, and 296, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 294, 326, and 324, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 94, and 89, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 94, and 99, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 230, 231, and 227, respectively.In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 261, and 259, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 274, 94, and 259, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 300, and 298, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 327, 328, and 324, respectively. In some embodiments, the HCDR associates with the LCDR to form an IBP4-binding region.

[0149] In some embodiments, the CDRs are determined according to Chothia numbering (see, e.g., Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). Thus, in some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 78, 79, and 80, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 78, 218, and 219, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 247, 248, and 249, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 247, 286, and 287, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 314, 315, and 316, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 90, 91, and 89, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 78, 79, and 80, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 228, 229, and 227, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 90, 260, and 259, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 272, 273, and 259, respectively.In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises an LCDR1, an LCDR2, and an LCDR3 set forth in SEQ ID NOs: 90, 299, and 298, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises an LCDR1, an LCDR2, and an LCDR3 set forth in SEQ ID NOs: 325, 326, and 324, respectively. In some embodiments, the HCDR associates with the LCDR to form an IBP4-binding region.

[0150] In some embodiments, the CDRs are determined according to AbM numbering. AbM hypervariable regions represent a compromise between Kabat CDRs and Chothia structural loops and are used by Oxford Moleculars AbM antibody modeling software (see, e.g., Martin, in Antibody Engineering, Vol. 2, Chapter 3, Springer Verlag). Thus, in some embodiments, an isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 81, 82, and 83, respectively. In some embodiments, an anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 81, 220, and 221, respectively. In some embodiments, an anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 250, 251, and 252, respectively. In some embodiments, the anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 288, 289, and 290, respectively. In some embodiments, the anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 288, 317, and 318, respectively. In some embodiments, the anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 94, and 95, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 94, and 100, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 230, 231, and 232, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 261, and 262, respectively.In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 274, 94, and 262, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 300, and 301, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 327, 328, and 329, respectively. In some embodiments, the HCDR associates with the LCDR to form an IBP4-binding region.

[0151] In some embodiments, the CDRs are determined according to Contact numbering. Thus, in some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 84, 85, and 86, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 84, 222, and 223, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 253, 254, and 255, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 291, 292, and 293, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 291, 319, and 320, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 96, 97, and 98, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 96, 97, and 101, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 233, 234, and 235, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 233, 234, and 235, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 96, 263, and 264, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 275, 97, and 264, respectively. In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 96, 302, and 298, respectively.In some embodiments, the isolated anti-IBP4 antibody or fragment thereof comprises LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 330, 331, and 332, respectively. In some embodiments, the HCDR associates with the LCDR to form an IBP4-binding region.

[0152] In some embodiments, the anti-IBP4 antibody is humanized. In some embodiments, the anti-IBP4 antibody is partially humanized. In some embodiments, the anti-IBP4 antibody is chimeric. In some embodiments, the anti-IBP4 antibody is a humanized chimeric. In some embodiments, the anti-IBP4 antibody is a murine antibody. In some embodiments, the anti-IBP4 antibody comprises an acceptor human framework, e.g., a human immunoglobulin framework or a human consensus framework. In some embodiments, the anti-IBP4 antibody comprises an acceptor mouse framework, e.g., a mouse immunoglobulin framework or a mouse consensus framework. Figures 21A-21B show exemplary heavy chain consensus CDRs and light chain consensus CDRs for anti-IBP4 antibodies, respectively.

[0153] In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 75-77, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 78-80, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 81-83, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 84-86, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 118, 119, and 83, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 75, 216, and 217, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 78, 218, and 219, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 81, 220, and 221, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 84, 222, and 223, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 118, 224, and 225, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 244, 245, and 246, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 247, 248, and 249, respectively.In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 250, 251, and 252, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 253, 254, and 255, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 256, 257, and 258, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 244, 284, and 285, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 247, 286, and 287, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 288, 289, and 290, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 291, 292, and 293, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 294, 295, and 296, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 244, 312, and 313, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 314, 315, and 316, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 288, 317, and 318, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 291, 319, and 320, respectively.In some embodiments, the anti-IBP4 antibodies provided herein comprise HCDR1, HCDR2, and HCDR3 as set forth in SEQ ID NOs: 294, 321, and 322, respectively. The CDR sequences can be determined according to well-known numbering systems or combinations thereof. In some embodiments, the CDRs are according to IMGT numbering. In some embodiments, the CDRs are according to AbM numbering. In other embodiments, the CDRs are according to Chothia numbering. In other embodiments, the CDRs are according to Contact numbering.

[0154] In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 87, 24, and 89, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 90, 91, and 89, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 94, and 95, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 96, 97, and 98, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 94, and 89, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 87, 24, and 99, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 90, 91, and 99, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 94, and 100, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 96, 97, and 101, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 94, and 99, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 226, 24, and 227, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 228, 229, and 227, respectively.In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 230, 231, and 232, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 233, 234, and 235, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 230, 231, and 227, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 87, 24, and 259, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 90, 260, and 259, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 261, and 262, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 96, 263, and 264, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 261, and 259, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 23, 24, and 259, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 272, 273, and 259, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 275, 94, and 262, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 275, 97, and 264, respectively.In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 274, 94, and 259, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 87, 297, and 298, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 90, 299, and 298, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 300, and 301, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 96, 302, and 303, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 300, and 298, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 323, 24, and 324, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 325, 326, and 324, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 327, 328, and 329, respectively. In some embodiments, the anti-IBP4 antibodies provided herein comprise LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 330, 331, and 332, respectively. In some embodiments, the anti-IBP4 antibody provided herein comprises LCDR1, LCDR2, and LCDR3 as set forth in SEQ ID NOs: 327, 328, and 324, respectively. The CDR sequences can be determined according to well-known numbering systems or combinations thereof. In some embodiments, the CDRs are numbered according to the IMGT numbering system.In some embodiments, the CDRs are according to AbM numbering, in other embodiments, the CDRs are according to Chothia numbering, in other embodiments, the CDRs are according to Contact numbering.

[0155] In some embodiments, the antibodies or fragments thereof provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 75, 76, and 77, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 87, 24, and 89. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 78, 79, and 80, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 90, 91, and 89. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 81, 82, and 83, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 94, and 95. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 84, 85, and 86, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 96, 97, and 98. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 118, 119, and 83, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 94, and 89. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 75, 216, and 217, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 226, 24, and 227. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 78, 218, and 219, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 228, 229, and 227.In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 81, 220, and 221, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 230, 231, and 232. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 84, 222, and 223, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 233, 234, and 235. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 118, 224, and 225, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 230, 231, and 227. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 244, 245, and 246, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 87, 24, and 259. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 247, 248, and 249, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 90, 260, and 259. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 250, 251, and 252, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 261, and 262. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 253, 254, and 255, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 96, 263, and 264.In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 256, 257, and 258, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 261, and 259. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 244, 245, and 246, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 23, 24, and 259. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 247, 248, and 249, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 272, 273, and 259. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 250, 252, and 252, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 274, 94, and 262. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 253, 254, and 255, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 275, 97, and 264. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 256, 257, and 258, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 274, 94, and 259. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 244, 284, and 285, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 87, 297, and 298.In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 247, 286, and 287, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 90, 299, and 298. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 288, 289, and 290, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 300, and 301. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 291, 292, and 293, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 96, 302, and 303. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 294, 295, and 296, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 300, and 298. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 244, 312, and 313, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 323, 24, and 324. In some embodiments, antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 314, 315, and 316, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 325, 326, and 324. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 288, 317, and 318, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 327, 328, and 329.In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 291, 319, and 320, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 330, 331, and 332. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 294, 321, and 322, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 327, 328, and 324. Thus, the CDR sequences can be determined according to well-known numbering systems or combinations thereof. In some embodiments, the CDRs are according to IMGT numbering. In some embodiments, the CDRs are according to AbM numbering. In other embodiments, the CDRs are according to Chothia numbering. In other embodiments, the CDRs are according to Contact numbering.

[0156] In some embodiments, the antibodies or fragments thereof provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 75, 76, and 77, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 87, 24, and 99. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 78, 79, and 80, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 90, 91, and 99. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 81, 82, and 83, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 94, and 100. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 84, 85, and 86, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 96, 97, and 101. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise HCDR1, HCDR2, and HCDR3 set forth in SEQ ID NOs: 118, 119, and 83, and LCDR1, LCDR2, and LCDR3 set forth in SEQ ID NOs: 93, 94, and 99. Thus, the CDR sequences can be determined according to well-known numbering systems or combinations thereof. In some embodiments, the CDRs are according to IMGT numbering. In some embodiments, the CDRs are according to AbM numbering. In other embodiments, the CDRs are according to Chothia numbering. In other embodiments, the CDRs are according to Contact numbering.

[0157] In some embodiments, an HCDR1 comprises an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 284, and 314; (ii) an HCDR1 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 284, and 314; 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321; (iii) an HCDR2 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 31 (iv) an HCDR3 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330; (v) an LCDR1 comprising an amino acid sequence having 4%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity; (v) an LCDR2 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331;and / or (vi) an LCDR3 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 89, 95, 98, 99, 100%, ...

[0158] In some embodiments, an HCDR1 comprises an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314; (ii) an HCDR1 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294, and 314; , 284, 286, 289, 292, 295, 312, 315, 317, 319, and 321; (iii) an HCDR2 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, and 316; 318, 320, 322; (iv) an HCDR3 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330; (v) an LCDR1 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326, 328, and 331;and / or (vi) an LCDR3 comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any of SEQ ID NOs: 89, 95, 98, 99, 100%, ...

[0159] In some embodiments, the antibody provided herein is humanized antibody.The framework region described herein is determined based on the boundary of CDR numbering system.In other words, when CDR is determined by Kabat, IMGT or Chothia for example, framework region is the amino acid residue around the CDR of variable region from N-terminus to C-terminus in the format: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. For example, FR1 is defined as the amino acid residues N-terminal to the CDR1 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; FR2 is defined as the amino acid residues between the CDR1 and CDR2 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; FR3 is defined as the amino acid residues between the CDR2 and CDR3 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; and FR4 is defined as the amino acid residues C-terminal to the CDR3 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system.

[0160] In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a VH sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to the sequence of SEQ ID NO: 107, and a VL sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 109. In some embodiments, an antibody or antigen-binding fragment provided herein comprises a VH sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 72 and a VL sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 74. In some embodiments, an antibody or antigen-binding fragment provided herein comprises a VH sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 213 and a VL sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 215. In some embodiments, an antibody or antigen-binding fragment provided herein comprises a VH sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 241 and a VL sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 243.In some embodiments, an antibody or antigen-binding fragment provided herein comprises a VH sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 241 and a VL sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 271. In some embodiments, an antibody or antigen-binding fragment provided herein comprises a VH sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 281 and a VL sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 283. In some embodiments, an antibody or antigen-binding fragment provided herein comprises a VH sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 309 and a VL sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to SEQ ID NO: 311. In further embodiments, one or more amino acid residues "I", "R", "S" or "V" corresponding to positions 58, 65, 116, and 117, respectively, in the linear sequence of SEQ ID NO: 309 are not present (e.g., are replaced by a different amino acid residue) in the VH sequence of an antibody or antigen-binding fragment provided herein.In still further embodiments, one or more amino acid residues "T", "A", "L", "N", "T", "M", "V", "S", or "S" corresponding to positions 14, 32, 46, 53, 56, 78, 87, 88, and 92, respectively, in the linear sequence of SEQ ID NO: 311 are not present (e.g., are replaced with a different amino acid residue) in the VL sequence of an antibody or antigen-binding fragment provided herein. In another embodiment, one or more amino acid residues "R", "T", "L", "E", "H", "T", "P", "V", "H", "G", "A", "D", "E", "T", "I", "R", "I", "A", "N", "Y", "I", "S", "Y", "A", "V", "S", or "V" corresponding to positions 13, 19, 20, 33, 35, 40, 41, 42, 43, 44, 50, 52, 54, 55, 58, 65, 69, 72, 99, 100, 102, 104, 106, 107, 108, 116, and 117, respectively, in the linear sequence of SEQ ID NO: 309 are not present (e.g., are replaced by a different amino acid residue) in the VH sequence of an antibody or antigen-binding fragment provided herein. In a further embodiment, one or more amino acid residues "T," "E," "I," "A," "L," "N," "T," "M," "D," "V," "S," or "S," corresponding to positions 14, 17, 21, 32, 46, 53, 56, 78, 85, 87, 88, and 92, respectively, in the linear sequence of SEQ ID NO: 311, are not present (e.g., are replaced with a different amino acid residue) in the VL sequence of an antibody or antigen-binding fragment provided herein. In another embodiment, one or more amino acid residues "I," "R," "A," "N," "Y," "I," "Y," "A," "V," "S," or "V," corresponding to positions 58, 65, 97, 99, 100, 102, 105, 107, 108, 116, and 117, respectively, in the linear sequence of SEQ ID NO: 309, are not present (e.g., are replaced with a different amino acid residue) in the VH sequence of an antibody or antigen-binding fragment provided herein.In further embodiments, one or more amino acid residues "T", "E", "I", "A", "L", "N", "T", "M", "V", "S", "S", "A", or "L" corresponding to positions 14, 17, 21, 32, 46, 53, 56, 78, 87, 88, 92, 100, and 106, respectively, in the linear sequence of SEQ ID NO: 311 are not present (e.g., are replaced with a different amino acid residue) in the VL sequence of an antibody or antigen-binding fragment provided herein.

[0161] In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a heavy chain sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to any one of SEQ ID NOs: 68, 103, 123, 128, 209, 237, 266, 277, 305, 333, 335, 337, 339, or 341. In some embodiments, the heavy or light chain comprises a C-terminal lysine residue (e.g., as shown underlined in SEQ ID NOs: 68, 103, 209, 237, 266, 277, and 305). In some embodiments, the C-terminal lysine residue is removed from the heavy chain. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a light chain sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any one of SEQ ID NOs: 70, 105, 122, 129, 211, 239, 268, 279, or 307. In some embodiments, the heavy or light chain comprises a C-terminal lysine residue (e.g., as underlined in SEQ ID NOs: 70 and 105). In some embodiments, the C-terminal lysine residue is removed from the light chain. In some embodiments, the heavy chain sequence associates with the light chain sequence to form an IBP4-binding region.

[0162] In certain embodiments, the antibodies or antigen-binding fragments thereof described herein comprise an amino acid sequence having a certain percent identity to any of the antibodies provided herein, for example, those set forth in the sequence listing below.

[0163] The percent identity between two sequences (for example, amino acid sequence or nucleic acid sequence) can be determined by mathematical algorithm.A non-limiting example of the mathematical algorithm used for comparing two sequences is the algorithm of Karlin and Altschul, Proc. Natl. Acad. Sci. USA 87:2264 2268 (1990), modified as in Karlin and Altschul, Proc. Natl. Acad. Sci. USA 90:5873 5877 (1993).Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul et al., J. Mol. Biol. 215:403 (1990).BLAST nucleotide search can be performed by setting the parameters of NBLAST nucleotide program, for example, score=100, word length=12, to obtain the nucleotide sequence homologous to the nucleic acid molecules described herein. BLAST protein searches can be performed by setting the parameters of the XBLAST program, for example, to score 50 and word length = 3, to obtain amino acid sequences homologous to the protein molecules described herein. To obtain gapped alignments for comparison purposes, Gapped BLAST can be used, as described in Altschul et al., Nucleic Acids Res. 25:3389 3402 (1997). Alternatively, PSI BLAST can be used to perform an iterated search that detects distant relationships between molecules (ibid.). When using BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of each program (e.g., XBLAST and NBLAST) can be used (see, for example, the National Center for Biotechnology Information (NCBI) on the World Wide Web at ncbi.nlm.nih.gov).Another non-limiting example of a mathematical algorithm utilized for comparing sequences is the algorithm of Myers and Miller, CABIOS 4:11-17 (1998). Such an algorithm is incorporated into the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package. When utilizing the ALIGN program to compare amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used. The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percent identity, typically, only perfect matches are counted.

[0164] In one aspect, the anti-IBP4 antibodies or fragments thereof disclosed herein may contain some variation in LCDR3. Thus, provided herein is an isolated anti-IBP4 antibody or fragment thereof comprising a heavy chain comprising an HCDR1 comprising the amino acid sequence GYTFTDYY (SEQ ID NO: 75), an HCDR2 comprising the amino acid sequence INPNNGGS (SEQ ID NO: 76), and an HCDR3 comprising the amino acid sequence ARWDGGFDY (SEQ ID NO: 77); and a light chain comprising an LCDR1 comprising the amino acid sequence QNVGTN (SEQ ID NO: 87), an LCDR2 comprising the amino acid sequence SAS (SEQ ID NO: 24), and an LCDR3 comprising the amino acid sequence QQYNX1YPLT (SEQ ID NO: 114). The residue identified as X1 can independently be any naturally occurring amino acid residue. For example, the residue identified as X1 can be isoleucine or serine. In some embodiments, X1 is isoleucine. In other embodiments, X1 is serine.

[0165] In another embodiment, the anti-IBP4 antibodies or fragments thereof disclosed herein may contain some variation in one or more of the HCDR1-HDCDR3 or LCDR1-LCDR3 sequences disclosed herein, such as (i) an HCDR1 comprising the amino acid sequence GYTFTYX1 (SEQ ID NO: 354), an HCDR2 comprising the amino acid sequence IX2X3X4X5GGX6 (SEQ ID NO: 355), and an HCDR3 comprising the amino acid sequence X7RX8X9X 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 DX 18 (ii) a heavy chain comprising an HCDR3 comprising the amino acid sequence QNVX 19 X 20 X 21 LCDR1 comprising (SEQ ID NO: 357), amino acid sequence SX 22 LCDR2 containing S (SEQ ID NO: 358), and the amino acid sequence QQYX 23 X 24 YPLX 25( Provided herein is an isolated anti-IBP4 antibody or fragment thereof comprising a light chain comprising an LCDR3 comprising SEQ ID NO: 359. In some embodiments, X1 to X5, X7, X8, X9, X10, X11, X12, X13, X14, X15, X16, X17, X18, X19, X20, X21, X22, X23, X24, X25, X26, X27, X 12 ~X 14 , X 18 ~X 20 , X 22 , and X 24 ~X 25 are independently any naturally occurring amino acid residue, X6 is independently any naturally occurring amino acid residue except isoleucine, X8 is independently a deletion or any naturally occurring amino acid residue except asparagine, X9 is independently any naturally occurring amino acid residue except tyrosine, and X 10 are independently a deletion or any naturally occurring amino acid residue, and X 11 are independently deleted, or any naturally occurring amino acid residue except isoleucine, and X 15 are independently a deletion or any naturally occurring amino acid residue, and X16 are independently a deletion or any naturally occurring amino acid residue except alanine, and X 17 are independently any naturally occurring amino acid residue except valine, and X 21 are independently any naturally occurring amino acid residue except alanine, and X 23 are independently any naturally occurring amino acid residue except serine. In some embodiments, X1 is tyrosine or glutamic acid. In some embodiments, X2 is asparagine or aspartic acid. In some embodiments, X3 is proline or threonine. In some embodiments, X4 is glutamic acid or asparagine. In some embodiments, X5 is threonine or asparagine. In some embodiments, X6 is alanine or proline, and X7 is alanine or threonine. In some embodiments, X8 is alanine. In some embodiments, X9 is tryptophan or arginine. In some embodiments, X 10 is glycine. 11 is tryptophan or serine. 12 is aspartic acid or arginine or serine. 13 is arginine or serine or glycine. 14 is tyrosine or glycine or valine. 15 is tyrosine. 16 is tyrosine. 17 is phenylalanine. 18 is tyrosine or asparagine. 19 is glycine or aspartic acid. 20 is threonine or isoleucine. 21 is aspartic acid or asparagine. 22is alanine or threonine. 23 is asparagine or aspartic acid. 24 is isoleucine or serine or threonine. 25 is alanine or threonine. In some embodiments, the anti-IBP4 antibody or fragment thereof disclosed herein comprises an HCDR1 comprising the amino acid sequence GYTFTYX1 (SEQ ID NO: 354), an HCDR2 comprising the amino acid sequence IX2X3X4X5GGX6 (SEQ ID NO: 355), and an HCDR3 comprising the amino acid sequence X7RX8X9X 10 X 11 X 12 X 13 X 14 X 15 X 16 X 17 DX 18 (ii) an HCDR3 comprising the amino acid sequence QNVX; and 19 X 20 X 21 LCDR1 comprising (SEQ ID NO: 357), amino acid sequence SX 22 LCDR2 containing S (SEQ ID NO: 358), and the amino acid sequence QQYX 23 X 24 YPLX 25 (SEQ ID NO: 359), wherein X1 to X 25 Any one of the positions has a combination of amino acid residues described herein. For example, for HCDR2, in some embodiments, X2 is asparagine or aspartic acid, X3 is proline or threonine, X4-X5 are any naturally occurring amino acid residues, and X6 is any naturally occurring amino acid residue except isoleucine. As another example, for LCDR1, in some embodiments, X 19 is any naturally occurring amino acid residue, and X 20 is threonine or isoleucine, and X 21 is any naturally occurring amino acid residue except alanine. 22is alanine or threonine. In yet another example, for LCDR3, in some embodiments, X 23 is any naturally occurring amino acid residue except serine, and X 24 is isoleucine, serine, or threonine, and X 25is alanine or threonine. Figures 21A-21B show exemplary heavy and light chain IMGT consensus CDRs (complementarity determining regions) for, e.g., anti-IBP4 antibodies 1H9.B6, 5C11.A7, 7G11.H4, 5B3.G9, 7D6.G4, 3D4.E9, and 1G3.A3. The nomenclature of the CDRs is shown below the sequences. Exemplary HCDRs and LCDRs and their respective SEQ ID NOs can be identified, e.g., in any of Tables 18, 20, 24, 26, 28, 30, and 32. Exemplary consensus CDR sequences for anti-SHBG and anti-IBP4 antibodies and their respective SEQ ID NOs can be identified, e.g., in Table 22. Figure 21A shows exemplary heavy chain IMGT consensus CDRs. The sequences for each CDRH1, CDRH2, and CDRH3 are shown in the left, center, and right columns, respectively (SEQ ID NOs: 275, 216, and 217 for 1H9.B6; SEQ ID NOs: 244, 245, and 246 for 5C11.A7 and 7G11.H4; SEQ ID NOs: 75, 216, and 77 for 5B3.G9 and 7D6.G4; SEQ ID NOs: 244, 284, and 285 for 3D4.E9; and SEQ ID NOs: 244, 312, and 313 for 1G3.A3). Figure 21B shows exemplary light chain IMGT consensus CDRs. The sequences for each CDRL1, CDRL2, and CDRL3 are shown in the left, center, and right columns, respectively (SEQ ID NOs: 226, 24, and 227 for 1H9.B6; SEQ ID NOs: 87, 24, and 259 for 5C11.A7; SEQ ID NOs: 23, 24, and 259 for 7G11.H4; SEQ ID NOs: 87, 24, and 89 for 5B3.G9; SEQ ID NOs: 87, 24, and 99 for 7D6.G4; SEQ ID NOs: 87, 297, and 298 for 3D4.E9; and SEQ ID NOs: 323, 24, and 324 for 1G3.A3). Figures 22A-22B show exemplary heavy and light chain variable regions for anti-IBP4 antibodies 1G3.A3, 1H9.B6, 3D4.E9, 5C11.A7, 7G11.H4, 7D6.G4, and 5B3.G9. Bold and underlined residues indicate less preferred residues from the 1G3.A3 clone according to the alignment of these antibodies.Exemplary heavy and light chain variable regions and their respective SEQ ID NOs can be identified, for example, in any of Tables 17, 19, 23, 25, 27, 29, and 31. Figure 22A shows the heavy chain variable regions for 1G3.A3 (SEQ ID NO: 309), 1H9.B6 (SEQ ID NO: 213), 3D4.E9 (SEQ ID NO: 281), 5C11.A7 (SEQ ID NO: 241), 7G11.H4 (SEQ ID NO: 241), 7D6.G4 (SEQ ID NO: 107), and 5B3.G9 (SEQ ID NO: 72). Figure 22B shows the light chain variable regions for 1G3.A3 (sequence number 311), 1H9.B6 (sequence number 215), 3D4.E9 (sequence number 283), 5C11.A7 (sequence number 243), 7G11.H4 (sequence number 271), 7D6.G4 (sequence number 109), and 5B3.G9 (sequence number 74).

[0166] In another aspect, provided herein is an isolated anti-IBP4 antibody or fragment thereof comprising: (i) a heavy chain comprising an HCDR1 comprising the amino acid sequence GYTFTDYY (SEQ ID NO: 75), an HCDR2 comprising the amino acid sequence INPNNGGX1 (SEQ ID NO: 360), and an HCDR3 comprising the amino acid sequence ARWX2X3DX4X5FDY (SEQ ID NO: 361); and (ii) a light chain comprising an LCDR1 comprising the amino acid sequence QNVGX6X7 (SEQ ID NO: 362), an LCDR2 comprising the amino acid sequence SAS (SEQ ID NO: 24), and an LCDR3 comprising the amino acid sequence QQYNX8YPLX9 (SEQ ID NO: 363). In some embodiments, X1 is independently any naturally occurring amino acid residue except isoleucine, X2 and X3 are each independently deleted or any naturally occurring amino acid residue, X4 is independently any naturally occurring amino acid residue except serine, X5, X6, X8, and X9 are each independently any naturally occurring amino acid residue, and X7 is independently any naturally occurring amino acid residue except alanine. In some embodiments, X1 is alanine or serine. In some embodiments, X2 is glycine. In some embodiments, X3 is tryptophan. In some embodiments, X4 is arginine or glycine. In some embodiments, X5 is tyrosine or glycine. In some embodiments, X6 is isoleucine or threonine. In some embodiments, X7 is aspartic acid or asparagine. In some embodiments, X8 is serine or isoleucine. In some embodiments, X9 is alanine or threonine.In some embodiments, the anti-IBP4 antibody or fragment thereof disclosed herein contains a light chain comprising: (i) an HCDR1 comprising the amino acid sequence GYTFTDYY (SEQ ID NO: 75), an HCDR2 comprising the amino acid sequence INPNNGGX1 (SEQ ID NO: 360), and an HCDR3 comprising the amino acid sequence ARWX2X3DX4X5FDY (SEQ ID NO: 361); and (ii) an LCDR1 comprising the amino acid sequence QNVGX6X7 (SEQ ID NO: 362), an LCDR2 comprising the amino acid sequence SAS (SEQ ID NO: 24), and an LCDR3 comprising the amino acid sequence QQYNX8YPLX9 (SEQ ID NO: 363), wherein any one of positions X1 through X9 has a combination of amino acid residues described herein. For example, for HCDR3, in some embodiments, X2 is deleted, X3 is tryptophan, X4 is arginine or glycine, and X5 is any naturally occurring amino acid residue. In another example, for LCDR1, in some embodiments, X6 is any naturally occurring amino acid residue and X7 is aspartic acid or asparagine. In another example, for LCDR3, in some embodiments, X8 is serine or isoleucine and X9 is any naturally occurring amino acid residue.

[0167] In another aspect, (i) an HCDR1 comprising the amino acid sequence GYTFTDYE (SEQ ID NO: 244), the amino acid sequence IDX1ETGGX 2(Provided herein is an isolated anti-IBP4 antibody or fragment thereof, comprising: (i) a heavy chain comprising an HCDR1 comprising the amino acid sequence QNVX5TN (SEQ ID NO: 364), an HCDR2 comprising the amino acid sequence TRARGSX3SVYYFDX4 (SEQ ID NO: 365), and an HCDR3 comprising the amino acid sequence QQYDX7YPLT (SEQ ID NO: 368); and (ii) a light chain comprising an LCDR1 comprising the amino acid sequence QNVX5TN (SEQ ID NO: 366), an LCDR2 comprising the amino acid sequence SX6S (SEQ ID NO: 367), and an LCDR3 comprising the amino acid sequence QQYDX7YPLT (SEQ ID NO: 368). In some embodiments, each of X1, X3, X4, X5, X6, and X7 is independently any naturally occurring amino acid residue, and X2 is independently any naturally occurring amino acid residue except isoleucine. In some embodiments, X1 is proline or threonine. In some embodiments, X2 is proline or alanine. In some embodiments, X3 is serine or arginine. In some embodiments, X4 is tyrosine or asparagine. In some embodiments, X5 is glycine or aspartic acid. In some embodiments, X6 is alanine or threonine. In some embodiments, X7 is serine or threonine. In some embodiments, the anti-IBP4 antibody or fragment thereof disclosed herein contains a light chain comprising (i) an HCDR1 comprising the amino acid sequence GYTFTDYE (SEQ ID NO: 244), an HCDR2 comprising the amino acid sequence IDX1ETGGX2 (SEQ ID NO: 364), and an HCDR3 comprising the amino acid sequence TRARGSX3SVYYFDX4 (SEQ ID NO: 365); and (ii) an LCDR1 comprising the amino acid sequence QNVX5TN (SEQ ID NO: 366), an LCDR2 comprising the amino acid sequence SX6S (SEQ ID NO: 367), and an LCDR3 comprising the amino acid sequence QQYDX7YPLT (SEQ ID NO: 368), wherein any one of positions X1 through X7 has a combination of amino acid residues described herein. For example, for HCDR2, in some embodiments, X1 is proline or threonine, and X2 is any naturally occurring amino acid residue. In another example, for HCDR3, in some embodiments, X3 is any naturally occurring amino acid residue and X4 is tyrosine or asparagine.In another example, for LCDR3, in some embodiments, X7 is serine.

[0168] Figures 23A-23B, for example, show exemplary CLUSTAL O(1.2.4) multiple sequence alignments for the heavy chain variable regions of two groups of anti-IBP4 antibodies: 1H9.B6, 7D6.G4, 5B3.G9, and 1G3.A3 (Group 1) in one group, and 3D4.E9, 5C11.A7, 7G11.H4, and 1G3.A3 (Group 2) in the other group. HCDRs are highlighted for each alignment and group. Bold and underlined residues indicate less favored residues from the 1G3.A3 clone according to the alignment of these antibodies. CDR nomenclature is shown below the sequence alignment for each group. Exemplary consensus CDR sequences for anti-SHBG and anti-IBP4 antibodies, as well as their respective SEQ ID NOs, can be identified, for example, in Table 22. Figure 23A shows an exemplary sequence alignment for the heavy chain variable regions of group 1: 1G3.A3 (SEQ ID NO: 309), 1H9.B6 (SEQ ID NO: 213), 7D6.G4 (SEQ ID NO: 107), and 5B3.G9 (SEQ ID NO: 72). Figure 23B shows an exemplary sequence alignment for the heavy chain variable regions of group 2: 1G3.A3 (SEQ ID NO: 309), 3D4.E9 (SEQ ID NO: 281), 5C11.A7 (SEQ ID NO: 241), and 7G11.H4 (SEQ ID NO: 241). Figures 24A-24B also show exemplary CLUSTAL O(1.2.4) multiple sequence alignments for the light chain variable regions of two groups of anti-IBP4 antibodies: 1H9.B6, 7D6.G4, 5B3.G9, and 1G3.A3 (group 1), and 3D4.E9, 5C11.A7, 7G11.H4, and 1G3.A3 (group 2). The LCDRs for each alignment and group are highlighted. Bold and underlined residues indicate less favored residues from the 1G3.A3 clone according to the alignment of these antibodies. The CDR nomenclature is shown below the sequence alignment for each group. Figure 24A shows an exemplary sequence alignment for the light chain variable regions of Group 1: 1G3.A3 (SEQ ID NO: 311), 1H9.B6 (SEQ ID NO: 215), 7D6.G4 (SEQ ID NO: 109), and 5B3.G9 (SEQ ID NO: 74).Figure 24B shows an exemplary sequence alignment for the light chain variable regions of Group 2: 1G3.A3 (SEQ ID NO: 311), 3D4.E9 (SEQ ID NO: 283), 5C11.A7 (SEQ ID NO: 243), and 7G11.H4 (SEQ ID NO: 271).

[0169] In some embodiments, the antibodies provided herein contain substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but the anti-IBP4 antibody comprising that sequence retains the ability to bind to IBP4. In some embodiments, a total of 1 to 10 amino acids are substituted, inserted, and / or deleted in the reference amino acid sequence. In some embodiments, the substitutions, insertions, or deletions occur in regions outside the CDRs (i.e., in the FRs). Optionally, the anti-IBP4 antibodies provided herein include post-translational modifications of the reference sequence.

[0170] In some embodiments, functional epitopes can be mapped, for example, by combinatorial alanine scanning, to identify amino acids in the IBP4 protein that are required for interaction with the anti-IBP4 antibodies provided herein. In some embodiments, the conformation and crystal structure of an anti-IBP4 antibody bound to IBP4 can be used to identify the epitope. In some embodiments, the present disclosure provides antibodies that specifically bind to the same epitope as any of the anti-IBP4 antibodies provided herein. For example, in some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 72 and a VL comprising the amino acid sequence of SEQ ID NO: 74. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 107 and a VL comprising the amino acid sequence of SEQ ID NO: 109. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 213 and a VL comprising the amino acid sequence of SEQ ID NO: 215. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 241 and a VL comprising the amino acid sequence of SEQ ID NO: 243. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 241 and a VL comprising the amino acid sequence of SEQ ID NO: 271. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 281 and a VL comprising the amino acid sequence of SEQ ID NO: 283. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 309 and a VL comprising the amino acid sequence of SEQ ID NO: 311.

[0171] In some embodiments, provided herein are anti-IBP4 antibodies or antigen-binding fragments thereof that specifically bind to IBP4 competitively with any one of the anti-IBP4 antibodies described herein. For example, in some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 72 and a VL comprising the amino acid sequence of SEQ ID NO: 74. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 107 and a VL comprising the amino acid sequence of SEQ ID NO: 109. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 213 and a VL comprising the amino acid sequence of SEQ ID NO: 215. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 241 and a VL comprising the amino acid sequence of SEQ ID NO: 243. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 241 and a VL comprising the amino acid sequence of SEQ ID NO: 271. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 281 and a VL comprising the amino acid sequence of SEQ ID NO: 283. In some embodiments, the antibodies or antigen-binding fragments provided herein bind to the same epitope as an anti-IBP4 antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 309 and a VL comprising the amino acid sequence of SEQ ID NO: 311.

[0172] In some embodiments, provided herein is an IBP4-binding agent comprising any one of the anti-IBP4 antibodies described above. In some embodiments, the IBP4-binding agent is a monoclonal antibody, including a murine antibody, a chimeric antibody, a humanized antibody, or a human antibody. In some embodiments, the anti-IBP4 antibody is an antibody fragment, e.g., an scFv. In some embodiments, the IBP4-binding agent is a fusion protein comprising the anti-IBP4 antibody provided herein. In other embodiments, the IBP4-binding agent is a multispecific antibody comprising the anti-IBP4 antibody provided herein. Other exemplary IBP4-binding agents are described in more detail in the following sections.

[0173] In some embodiments, an anti-IBP4 antibody or antigen binding protein according to any of the above embodiments may incorporate any single or combination of features, as described below.

[0174] In another aspect, the present disclosure relates to nucleic acid molecules encoding the heavy and / or light chains of the antibodies or fragments thereof disclosed herein. The nucleic acid molecules are preferably isolated nucleic acid molecules. Isolated nucleic acid molecules can be substantially pure nucleic acid molecules. In some embodiments, the nucleic acid molecules encode the heavy chain of an anti-IBP4 antibody or fragment thereof. In some embodiments, the nucleic acid molecule encoding the heavy chain has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 67, 102, 204, 206, 208, 236, 265, 276, 304, 334, 336, 338, 340, or 342. In some embodiments, the nucleic acid molecules encode the light chain of an anti-IBP4 antibody or fragment thereof. In some embodiments, the nucleic acid molecule encoding the light chain has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 69, 104, 205, 207, 210, 238, 267, 278, or 306. Methods and compositions for producing antibodies

[0175] Antibodies or fragments thereof useful in the present methods, compositions, kits, etc. can be produced, for example, by recombinant expression techniques from hybridomas, myelomas, or other antibody-expressing mammalian cells. As is well recognized in the art, recombinant expression of an antibody or fragment thereof that binds to a target antigen typically involves constructing an expression vector containing nucleic acid encoding the polypeptides (e.g., heavy and light chains) that form the antibody or fragment. Once nucleic acid encoding such a polypeptide is obtained, vectors for production of the antibody can be produced by recombinant DNA technology using techniques well known in the art. For example, for recombinant expression of an antibody, an expression vector can encode its heavy or light chain, or heavy or light chain variable domains, operably linked to a promoter. The expression vector can contain, for example, nucleotide sequences encoding the constant region of an antibody molecule, and the variable domain of the antibody can be cloned into such a vector for expression of the entire heavy or light chain. The expression vector can be transferred into a host cell by techniques known in the art.

[0176] In particular, techniques for producing single-chain antibodies can also be used in accordance with the methods disclosed herein. Single-chain antibodies (e.g., scFvs or single-chain variable fragments) comprise heavy and light chain variable regions connected with a linker peptide. In scFvs, the heavy and light chain variable regions can be derived from the same or different antibodies. ScFvs can be synthesized using recombinant techniques, for example, by expressing a vector encoding an scFv in a host organism such as E. coli. DNA encoding an scFv can be obtained by amplifying the entire DNA or a partial DNA encoding the desired amino acid sequence selected from DNA encoding the heavy chain or the heavy chain variable region of the above-mentioned antibody and DNA encoding the light chain or the light chain variable region as a template using PCR with primer pairs defining both ends of the DNA, and further amplifying DNA encoding a polypeptide linker portion and primer pairs defining both ends thereof, thereby ligating both ends of the linker to the heavy chain and light chain, respectively. An expression vector containing DNA encoding an scFv and a host transformed with the expression vector can be obtained according to conventional methods known in the art.

[0177] In one aspect, the present disclosure provides an antibody or fragment thereof produced by any of the methods disclosed herein.In some embodiments, the antibody or fragment thereof produced by any of the methods disclosed herein is the isolated anti-IBP4 antibody or fragment thereof described herein.For example, using the methods disclosed herein, (i) HCDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 75, 78, 81, 84, 118, 244, 247, 250, 253, 256, 288, 291, 294 and 314; and SEQ ID NO: 76, 79, 82, 85, 119, 216, 218, 220, 222, 224, 245, 248, 251, 254, 257, 284, HCDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 77, 80, 83, 86, 217, 219, 221, 223, 225, 246, 249, 252, 255, 258, 285, 287, 290, 293, 296, 313, 316, 318, 320, 322 or (ii) an LCDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23, 87, 90, 93, 96, 226, 228, 230, 233, 272, 274, 275, 323, 325, 327, and 330; and SEQ ID NOs: 24, 91, 94, 97, 229, 231, 234, 260, 261, 263, 273, 297, 299, 300, 302, 326 , 328, and 331; and a light chain comprising an LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 89, 95, 98, 99, 100, 101, 227, 232, 235, 259, 262, 264, 298, 301, 303, 324, 329, and 332. In some embodiments, the present disclosure provides a nucleic acid molecule encoding the heavy chain of an anti-IBP4 antibody or fragment thereof.In some embodiments, a nucleic acid molecule encoding a heavy chain has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 67, 102, 204, 206, 208, 236, 265, 276, 304, 334, 336, 338, 340, or 342. In some embodiments, the present disclosure provides a nucleic acid molecule encoding a light chain of an anti-IBP4 antibody or fragment thereof. In some embodiments, a nucleic acid molecule encoding a light chain has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 69, 104, 205, 207, 210, 238, 267, 278, or 306. In further embodiments, the present disclosure provides nucleic acid molecules encoding the heavy and light chains of an anti-IBP4 antibody or fragment thereof. In another aspect, the present disclosure provides a vector comprising a nucleic acid molecule encoding the heavy or light chain of an anti-IBP4 antibody or fragment thereof. In another aspect, the present disclosure provides an expression vector comprising a nucleic acid molecule encoding the heavy or light chain of an anti-IBP4 antibody or fragment thereof. In another aspect, the present disclosure provides a cell comprising any of the disclosed nucleic acid molecules, vectors, or expression vectors for the anti-IBP4 antibodies herein.

[0178] In some embodiments, the antibody or fragment thereof produced by any of the methods disclosed herein is an isolated SHBG antibody or fragment thereof described herein. For example, the methods disclosed herein can be used to produce an isolated anti-SHBG antibody or fragment thereof comprising: (i) a heavy chain comprising an HCDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 11, 14, 17, 20, 52, and 115; and an HCDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 12, 15, 18, 21, 44, 47, 50, 53, 116, and 117; and an HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 13, 16, 19, 22, 45, 48, 51, and 54; or (ii) a light chain comprising an LCDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23, 26, 29, and 32; and an LCDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 24, 27, 30, and 33; and an LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 25, 31, and 34. In some embodiments, the present disclosure provides a nucleic acid molecule encoding the heavy chain of an anti-SHBG antibody or fragment thereof. In some embodiments, the nucleic acid molecule encoding the heavy chain has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 3, 35, 200, or 202. In some embodiments, the present disclosure provides a nucleic acid molecule encoding the light chain of an anti-SHBG antibody, or fragment thereof. In some embodiments, the nucleic acid molecule encoding the light chain has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 5, 37, 201, or 203. In further embodiments, the present disclosure provides nucleic acid molecules encoding the heavy and light chains of an anti-SHBG antibody, or fragment thereof. In another aspect, the present disclosure provides a vector comprising a nucleic acid molecule encoding the heavy or light chain of an anti-SHBG antibody, or fragment thereof. In another aspect, the present disclosure provides an expression vector comprising a nucleic acid molecule encoding the heavy or light chain of an anti-SHBG antibody, or fragment thereof. In another aspect, the disclosure provides a cell comprising any of the disclosed nucleic acid molecules, vectors, or expression vectors for the anti-SHBG antibodies herein.

[0179] In some embodiments, the present disclosure provides a vector or expression vector comprising a nucleic acid molecule encoding the heavy chain of any of the anti-SHBG and anti-IBP4 antibodies or fragments thereof disclosed herein. In some embodiments, the vector or expression vector comprises a nucleic acid molecule encoding the light chain of any of the anti-SHBG and anti-IBP4 antibodies or fragments thereof disclosed herein. In some embodiments, the present disclosure provides a cell comprising any of the nucleic acid molecules, vectors, or expression vectors disclosed herein.

[0180] In some embodiments, an anti-SHBG or anti-IBP4 antibody or a fragment thereof can be a humanized antibody. A humanized antibody is an antibody molecule that binds to a desired antigen and has one or more complementarity-determining regions (CDRs) from a non-human species and a framework and constant region from a human immunoglobulin molecule. Often, framework residues in the human framework region are replaced with corresponding residues from the CDR donor antibody to alter or preferably improve antigen binding. As is well recognized in the art, various methods known in the art can be used to generate humanized antibodies having the characteristics of the anti-SHBG or anti-IBP4 antibody or fragment thereof described herein.

[0181] In some embodiments, anti-SHBG or anti-IBP4 antibody is human antibody.As is well recognized in the art, human antibody can be produced by various methods known in the art, such as phage display method using antibody library derived from human immunoglobulin sequence.Therefore, various methods known in the art can be used to generate human antibody with the characteristics of anti-SHBG or anti-IBP4 antibody or its fragment described herein. composition

[0182] In another aspect, the present disclosure relates to a composition comprising an isolated antibody or antigen-binding fragment thereof described herein and a carrier (e.g., a pharmaceutically acceptable carrier). In some embodiments, the antibody or fragment thereof comprises an anti-SHBG antibody or fragment thereof. In some embodiments, the anti-SHBG antibody or fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 11-22, 44, 45, 47, 48, 50-54, 115, and 117, and the light chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-34. In some embodiments, the heavy chain associates with the light chain to form a binding site for SHBG. In another aspect, the present disclosure relates to a composition comprising an isolated anti-IBP4 antibody or antigen-binding fragment thereof described herein and a carrier (e.g., a pharmaceutically acceptable carrier). In some embodiments, the anti-IBP4 antibody or antigen-binding fragment thereof comprises a heavy chain and / or a light chain, wherein the heavy chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 75-86, 118, 119, 216-225, 244-258, 284-296, and 312-322, and the light chain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 23, 24, 87-101, 226-235, 259-264, 272-275, 297-303, and 323-332. In some embodiments, the heavy chain associates with the light chain to form a binding site for IBP4.

[0183] In some embodiments, the compositions of the present disclosure comprise an anti-SHBG antibody or fragment thereof, wherein the heavy chain comprises a heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 11-22, 115, or 116, and a light chain comprising a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-34. In some embodiments, the heavy chain associates with the light chain to form a binding site for SHBG. In other embodiments, the anti-SHBG antibody or fragment thereof comprises a heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 11, 14, 17, 44, 45, 47, 48, and 50-54, and / or a light chain comprising a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-34. In some embodiments, the heavy chain associates with the light chain to form a binding site for SHBG.

[0184] In some embodiments, a composition of the present disclosure comprises an anti-IBP4 antibody or fragment thereof comprising a heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 75-86, 118, 119, 216-225, 244-258, 284-296, and 312-322, and a light chain comprising a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23, 87, 24, 89, 90, 91, 93, 94, 95-98, 226-235, 259-264, 272-275, 297-303, and 323-332. In some embodiments, the heavy chain associates with the light chain to form a binding site for IBP4. In other embodiments, the anti-IBP4 antibody or fragment thereof comprises a heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 75-86, 118, 119, 216-225, 244-258, 284-296, and 312-322, and / or a light chain comprising a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 87, 24, 90, 91, 93, 94, 96, 97, 99, 100, 101, 226-235, 259-264, 272-275, 297-303, and 323-332. In some embodiments, the heavy chain associates with the light chain to form a binding site for IBP4.

[0185] In some embodiments, the composition may comprise at least 0.1% by weight of an anti-SHBG antibody or antigen-binding fragment thereof. In some embodiments, the composition may comprise at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or more by weight of an anti-SHBG antibody or antigen-binding fragment thereof. In some embodiments, the composition may comprise at least 0.1% by weight of an anti-IBP4 antibody or antigen-binding fragment thereof. In some embodiments, the composition may have at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or more by weight of an anti-IBP4 antibody or antigen-binding fragment thereof. In some embodiments, the composition may have at least 0.1% by weight of an anti-SHBG antibody or antigen-binding fragment thereof in combination with an anti-IBP4 antibody or antigen-binding fragment thereof. In some embodiments, the composition can have at least 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or more by weight of an anti-SHBG antibody or antigen-binding fragment thereof and an anti-IBP4 antibody or antigen-binding fragment thereof.

[0186] In some embodiments, the composition is designed for use in an assay to detect SHBG and / or IBP4 from a sample. Suitable samples include, for example, blood, plasma, serum, amniotic fluid, vaginal secretions, saliva, and urine. In some embodiments, the biological sample is selected from the group consisting of whole blood, plasma, and serum. In certain embodiments, the biological sample is serum. In some embodiments, the assay can be used to detect SHBG and / or IBP4 in a sample from a pregnant woman. As discussed herein, detection of SHBG and / or IBP4 can indicate a woman's health status, such as the probability of preterm birth or preeclampsia.

[0187] In some embodiments, the composition is adapted for use in an immunoprecipitation assay, such as a co-immunoprecipitation assay. The assay can be used to detect the presence or absence of SHBG and / or anti-IBP4 antibodies in a sample from a pregnant woman. In some embodiments, the composition of the present disclosure includes an antibody capture agent that specifically binds to SHBG or IBP4 protein. The capture agent can include any one or more of the anti-SHBG and / or anti-IBP4 antibodies or fragments thereof described herein. Such capture agents can be prepared using any suitable method known in the art. See, for example, Coligan, Current Protocols in Immunology (1991); Harlow & Lane, Antibodies: A Laboratory Manual (1988); Goding, Monoclonal Antibodies: Principles and Practice (2d ed. 1986). The antibody capture agent can include any antibody or fragment thereof described herein, whether naturally occurring or wholly or partially synthetically produced. In some embodiments, the capture agent is linked to a solid support such as a bead, eg, a magnetic bead.

[0188] For example, in some embodiments, the compositions of the present disclosure comprise one or more anti-SHBG antibodies or fragments thereof and / or one or more anti-IBP4 antibodies or fragments thereof linked to a solid support, which can be used to capture, isolate, and / or detect one or more SHBG proteins or fragments thereof and / or IBP4 proteins or fragments thereof from a sample. The solid support can comprise any substrate. Exemplary substrates include antibody-coated 96-well plates or nitrocellulose membranes, which are subsequently probed for the presence of SHBG and / or IBP4 proteins. Alternatively, substrates that can comprise microspheres, microparticles, microbeads, beads, or other particles can be used for capturing and detecting SHBG and / or IBP4. In some embodiments, the compositions of the present disclosure comprise one or more anti-IBP4 antibodies or fragments thereof linked to a solid support, which can be used to capture and isolate one or more IBP4 proteins or fragments thereof from a sample.

[0189] For example, in some embodiments, the substrate is a bead. The bead can be covalently linked to a polypeptide that binds to SHBG and / or IBP4 protein. The polypeptide can include a sequence disclosed in the sequence listing below. The bead can be used in an assay to detect or measure the presence or absence of SHBG and / or IBP4 in a biological sample. For example, the bead can be deposited in a sample under conditions sufficient for the polypeptide to bind to SHBG and / or IBP4. After binding, the bead can be recovered from the sample, for example, using a magnet. Any SHBG and / or IBP4 present in the sample can be detected or measured by identifying the SHBG and / or IBP4 bound to the recovered beads. SHBG and / or IBP4 can be identified, for example, by mass spectrometry.

[0190] Thus, the compositions described herein may be useful for any existing, available, or conventional separation, detection, and quantification method that can be used to measure the presence or absence (e.g., the readout is presence vs. absence; or detectable amount vs. undetectable amount) and / or amount (e.g., the readout is absolute amount or relative amount, e.g., absolute concentration or relative concentration, etc.) of SHBG or IBP4 protein, peptide, polypeptide, protein, and / or fragment thereof, and, if desired, one or more other biomarkers or fragments thereof in a sample.

[0191] In some embodiments, the detection and / or quantification of SHBG or IBP4 comprises assays including enzyme immunoassays (EIAs), enzyme-linked immunosorbent assays (ELISAs), and radioimmunoassays (RIAs). In some embodiments, the detection and / or quantification of one or more of SHBG and IBP4 further comprises mass spectrometry (MS). In yet further embodiments, the mass spectrometry is co-immunoprecipitation-mass spectrometry (co-IP MS), a technique suitable for isolating the entire protein complex followed by analysis by mass spectrometry.

[0192] In some embodiments, the anti-SHBG and anti-IBP4 antibodies described herein include a carrier. The carrier can be any type of material, including soluble and insoluble materials, as long as it is useful for carrying the anti-SHBG and / or anti-IBP4 antibodies in an assay or during therapeutic treatment. For example, examples include beads, e.g., magnetic beads, and particles. The material and size of the carrier can be selected as needed depending on the type of assay or therapeutic use. In some embodiments, the carrier is a pharmaceutically acceptable carrier, including liquid, semi-solid, i.e., paste, or solid carriers. Examples of carriers or diluents include fats, oils, water, saline, lipids, liposomes, resins, binders, fillers, etc., or combinations thereof. Pharmaceutically acceptable carriers, as known to those skilled in the art, include aqueous solvents (e.g., water, alcoholic / aqueous solutions, ethanol, saline, parenteral vehicles such as sodium chloride, Ringer's dextrose, and the like), non-aqueous solvents (e.g., propylene glycol, polyethylene glycol, vegetable oils, and injectable organic esters such as ethyl oleate), dispersion media, coatings (e.g., lecithin), surfactants, antioxidants, preservatives (e.g., antibacterial or antifungal agents, antioxidants, chelating agents, inert gases, parabens (e.g., methylparaben, propylparaben), chloroform, ... Examples of suitable media, agents, diluents, or carriers include but are not limited to: sorbic acid, propyl alcohol, propyl butanol, phenol, sorbic acid, thimerosal), isotonic agents (e.g., sugars, sodium chloride), absorption delaying agents (e.g., aluminum monostearate, gelatin), salts, drugs, drug stabilizers (e.g., buffers, amino acids such as glycine and lysine, carbohydrates such as dextrose, mannose, galactose, fructose, lactose, sucrose, maltose, sorbitol, mannitol, etc.), gels, binders, excipients, disintegrants, lubricants, sweeteners, flavoring agents, dyes, fluid and nutritional supplements, similar materials, and combinations thereof. Except insofar as any conventional vehicle, agent, diluent, or carrier is deleterious to the recipient or to the therapeutic effectiveness of the composition contained therein, its use in administrable compositions for use in carrying out the methods is appropriate.The pH and exact concentrations of the various components in the pharmaceutical composition are adjusted according to well-known parameters. According to certain embodiments of the present disclosure, the composition can be combined with the carrier in any conventional and practical manner, i.e., by dissolving, suspending, emulsifying, mixing, encapsulating, absorbing, grinding, etc. Such procedures are routine for those skilled in the art.

[0193] In some embodiments, pharmaceutically acceptable carrier can be aqueous pH buffer solution.Examples include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight (for example, less than about 10 amino acid residues) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulin; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as TWEEN®, polyethylene glycol (PEG), and PLURONICS®.

[0194] In some embodiments, pharmaceutically acceptable carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Water can be a carrier, particularly when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, polysorbate-80, and the like. The compositions can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. These compositions can take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release formulations, and the like.

[0195] Certain embodiments of the present disclosure can have different types of carriers depending on whether they are administered in solid, liquid, or aerosol form and whether they need to be sterile for routes of administration such as injection. The compositions can be formulated for administration intravenously, intradermally, transdermally, intrathecally, intraarterially, intraperitoneally, intranasally, intravaginally, intrarectally, intramuscularly, subcutaneously, mucosally, orally, topically, locally, by inhalation (e.g., aerosol inhalation), by injection, by infusion, by continuous infusion, by local perfusion directly immersing target cells, via a catheter, via lavage, in lipid compositions (e.g., liposomes), or by other methods known to those skilled in the art or any combination of the foregoing (see, for example, Remington's Pharmaceutical Sciences, 18th Ed., 1990, incorporated herein by reference). Typically, such compositions can be prepared either as liquid solutions or suspensions; solid forms suitable for use in preparing solutions or suspensions by adding liquid prior to injection can also be prepared; the preparations can also be emulsified.

[0196] Anti-SHBG antibodies or other molecules having antigen-binding fragments that specifically bind to SHBG can be formulated into compositions in free base, neutral, or salt form. Similarly, anti-IBP4 antibodies or other molecules having antigen-binding fragments that specifically bind to IBP4 can be formulated into compositions in free base, neutral, or salt form. Pharmaceutically acceptable salts include acid addition salts, such as those formed with free amino groups of proteinaceous compositions, or with inorganic acids such as hydrochloric acid or phosphoric acid, or organic acids such as acetic acid, oxalic acid, tartaric acid, or mandelic acid. Salts formed with free carboxyl groups can also be derived from inorganic bases such as sodium, potassium, ammonium, calcium, or ferric hydroxide; or organic bases such as isopropylamine, trimethylamine, 2-ethylaminoethanol, histidine, or procaine.

[0197] In a further embodiment, a pharmaceutical composition comprising a lipid is provided herein. Lipids can broadly encompass a class of substances that are characteristically insoluble in water and extractable with organic solvents. Examples include compounds containing long-chain aliphatic hydrocarbons and their derivatives. Lipids can be naturally occurring or synthetic (i.e., engineered or produced by humans). Lipids can be biological substances. Biological lipids are well known in the art and include, for example, neutral lipids, phospholipids, phosphoglycerides, steroids, terpenes, lysolipids, glycosphingolipids, glycolipids, sulfatides, lipids with ether- and ester-linked fatty acids, polymerizable lipids, and combinations thereof. Compounds that are recognized by those skilled in the art as lipids, other than those specifically described herein, can also be used.

[0198] Those skilled in the art will be familiar with a range of techniques that can be used to disperse compositions in lipid vehicles.For example, antibody can be dispersed in a solution that contains lipid, dissolved in lipid, emulsified in lipid, mixed with lipid, combined with lipid, covalently bonded to lipid, contained as a suspension in lipid, contained or complexed with micelles or liposomes, or otherwise associated with lipid or lipid structure by any means known to those skilled in the art.Dispersion may or may not result in the formation of liposomes.

[0199] As those skilled in the art will understand, the compositions described herein are not limited by the specific nature of the therapeutic preparation. For example, such compositions can be provided in formulations with physiologically acceptable liquid, gel, or solid carriers, diluents, and excipients. These therapeutic preparations can be administered to mammals in a manner similar to other therapeutic agents, for example, for veterinary use using domestic animals and clinical use in humans. Generally, the dosage required for therapeutic effect varies according to the type of use and mode of administration, as well as the specific requirements of the individual subject. The actual dosage of the composition administered to an animal patient, including a human patient, can be determined by physical and physiological factors, such as body weight, severity of the condition, the type of disease being treated, previous or concurrent therapeutic interventions, the patient's specific symptoms, and the route of administration. Depending on the dosage and route of administration, the preferred dosage and / or the frequency of administration of an effective amount can vary according to the subject's response. In any event, the practitioner responsible for administration will determine the concentration of the active ingredient(s) in the composition and the appropriate dosage for the individual subject. kit

[0200] In another aspect, the present disclosure relates to a kit.In some embodiments, the kit comprises an isolated anti-SHBG antibody or fragment thereof described herein, one or more auxiliary reagents, a solid support, and packaging therefor.In some embodiments, the kit comprises an isolated anti-IBP4 antibody or fragment thereof, one or more auxiliary reagents, a solid support, and packaging therefor.In some embodiments, the kit comprises an isolated anti-SHBG antibody or fragment thereof, and an isolated anti-IBP4 antibody or fragment thereof, one or more auxiliary reagents, a solid support, and packaging therefor.

[0201] In some embodiments, the anti-SHBG antibody or fragment thereof comprises a heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 11-22, 115, or 116, and a light chain comprising a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-34. In some embodiments, the heavy chain associates with the light chain to form a binding site for SHBG. In other embodiments, the anti-SHBG antibody or fragment thereof comprises a heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 11, 14, 17, 44, 45, 47, 48, and 50-54, and / or a light chain comprising a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-34. In some embodiments, the heavy chain associates with the light chain to form a binding site for SHBG.

[0202] In some embodiments, the anti-IBP4 antibody or fragment thereof comprises a heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 75-86, 118, 119, 216-225, 244-258, 284-296, and 312-322, and a light chain comprising a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 87, 24, 89, 90, 91, 93, 94, 95-98, 226-235, 259-264, 272-275, 297-303, and 323-332. In some embodiments, the heavy chain associates with the light chain to form a binding site for IBP4. In other embodiments, the anti-IBP4 antibody or fragment thereof comprises a heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 75-86, 118, 119, 216-225, 244-258, 284-296, and 312-322, and / or a light chain comprising a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 24, 87, 90, 91, 93, 94, 96, 97, 99, 100, 101, 226-235, 259-264, 272-275, 297-303, and 323-332. In some embodiments, the heavy chain associates with the light chain to form a binding site for IBP4.

[0203] In some embodiments, the one or more auxiliary reagents are selected from the group consisting of a secondary antibody, an incubation buffer, an immobilization buffer, a washing buffer, a blocking buffer, a detection buffer, a detection reagent, and a detection instrument, or any combination thereof. In some embodiments, the detection probe comprises a label. In some embodiments, the label is selected from the group consisting of a fluorophore, an enzyme, a chemiluminescent moiety, a radioactive moiety, an organic dye, and a small molecule.

[0204] The secondary antibody may include, for example, an anti-human IgA antibody, an anti-human IgD antibody, an anti-human IgE antibody, an anti-human IgG antibody, or an anti-human IgM antibody. In some embodiments, the secondary antibody is an anti-bovine antibody. The secondary detection antibody may be a monoclonal or polyclonal antibody. The secondary antibody may be derived from any mammalian organism, including mouse, rat, hamster, goat, camel, chicken, rabbit, etc. The secondary antibody may be conjugated to an enzyme (e.g., horseradish peroxidase (HRP), alkaline phosphatase (AP), luciferase, etc.) or a dye (e.g., a colorimetric dye, a fluorescent dye, a fluorescence resonance energy transfer (FRET) dye, a time-resolved (TR)-FRET dye, etc.). In some embodiments, the secondary antibody is a polyclonal rabbit anti-human IgG antibody conjugated with HRP.

[0205] In some embodiments, the detection buffer can contain a fluorescent detection reagent or a luminescent detection reagent. In some other aspects, the luminescent detection reagent contains luminol or luciferin. Any detection buffer known in the art can be included in the kits of the present disclosure. In some embodiments, the detection buffer is a citrate-phosphate buffer (e.g., about pH 4.2).

[0206] In some embodiments, the detection reagent is a colorimetric detection reagent, a fluorescent detection reagent, or a chemiluminescent detection reagent. In some embodiments, the colorimetric detection reagent includes PNPP (p-nitrophenyl phosphate), ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)), or OPD (o-phenylenediamine). In some embodiments, the fluorescent detection reagent includes QuantaBlu™ or QuantaRed™ (Thermo Scientific, Waltham, MA). In some embodiments, the luminescent detection reagent includes luminol or luciferin. In some embodiments, the detection reagent includes a trigger (e.g., HO) and a tracer (e.g., an isoluminol conjugate).

[0207] In some embodiments, the wash buffer can include those known in the art, such as a tris(hydroxymethyl)aminomethane (Tris)-based buffer (e.g., Tris-buffered saline, TBS) or a phosphate buffer (e.g., phosphate-buffered saline, PBS). The wash buffer can include a detergent, such as an ionic or non-ionic detergent. In some embodiments, the wash buffer is a PBS buffer (e.g., about pH 7.4) containing Tween® 20 (e.g., about 0.05% Tween® 20).

[0208] In some embodiments, the solid support can include any support known in the art that can immobilize the proteins of the present disclosure. In some embodiments, the solid support is selected from the group consisting of a microtiter well plate, a slide (e.g., a glass slide), a chip (e.g., a protein chip, a biosensor chip, e.g., a Biacore chip), a microfluidic cartridge, a cuvette, a bead (e.g., a magnetic bead), a resin, a bead, a sphere, a particle, a membrane, a plate, and a test tube. In some embodiments, the bead, sphere, or particle has a diameter of micrometers or nanometers. In some embodiments, the membrane is selected from the group consisting of nitrocellulose, nylon, polyvinylidene fluoride (PVDF), and polyvinylidene difluoride. In some embodiments, the anti-SHBG antibody or antigen-binding fragment is conjugated to a solid support.

[0209] In some embodiments, the packaging in the kits described herein can include a label indicating that the kit is used for the diagnosis, prognosis, or monitoring of preterm birth or preeclampsia. In some embodiments, the kit packaging includes an FDA-approved label. The FDA-approved label can include a notice of FDA-approved use and instructions. In some embodiments, the kit packaging can include suitable container means, which is a container that does not react with the components of the kit. In some embodiments, the container means includes one or more sealed vials, such as Eppendorf tubes, containing the anti-SHBG antibodies or antigen-binding fragments described herein. The containers can be made of a sterilizable material, such as plastic or glass.

[0210] The kits provided herein can be adapted to specific assay techniques. In some embodiments, the kits are ELISA kits, Dot Blot kits, chemiluminescent immunoassay (CIA) kits, or multiplex kits. In some embodiments, an ELSA kit can include a wash buffer, a sample diluent, a secondary antibody-enzyme conjugate, a detection reagent, and a stop solution. In some embodiments, a Dot Blot kit includes a wash buffer, a sample diluent, a secondary antibody-enzyme conjugate, a detection reagent, and a stop solution. In some embodiments, a CIA kit includes a wash buffer, a sample diluent, a tracer (e.g., an isoluminol conjugate), and a trigger (e.g., H2O2). In some embodiments, a multiplex kit includes a wash buffer, a sample diluent, and a secondary antibody-enzyme conjugate.

[0211] In some embodiments, the kit includes a positive control. In some embodiments, the positive control is a sample containing SHBG. In some embodiments, the positive control is a sample containing IBP4. In some other embodiments, the kit provided herein includes instructions for using a subunit of the kit to detect SHBG or glycosylated SHBG, or IBP4 in a sample from a subject.

[0212] Also provided herein is a kit that can be used as a companion diagnostic for the probability of preterm birth and preeclampsia.In some embodiments, the kit can be used to select patients for specific intervention therapy designed to reduce the probability of preterm birth and preeclampsia.Intervention therapy includes but is not limited to those known in the art, including the administration of progesterone. How to use

[0213] According to one aspect, the present disclosure provides a method for detecting the presence or absence of one or more biomarkers from a sample. In some embodiments, the one or more biomarkers include SHBG. In some embodiments, the one or more biomarkers include IBP4. In some embodiments, the one or more biomarkers include SHBG and IBP4. For example, in some embodiments, the present disclosure provides a method comprising contacting a biological sample containing one or more biomarkers with an anti-SHBG antibody or fragment thereof and / or an anti-IBP4 antibody or fragment thereof described herein. The method further comprises detecting binding of the antibody or fragment thereof to the one or more biomarkers. Detecting binding may comprise, for example, a proteomics workflow including mass spectrometry quantification. In some embodiments, the detected biomarker is SHBG protein. In some embodiments, the detected biomarker is glycosylated SHBG protein. In some embodiments, the detected biomarker is unglycosylated SHBG protein. In some embodiments, the detected biomarker is IBP4 protein. In some embodiments, the biological sample is blood or serum. In some embodiments, the detecting comprises separating from the biological sample one or more biomarkers bound to the antibody or antibody fragment thereof, hi some embodiments, the detecting comprises an immunoassay selected from the group consisting of sandwich immunoassay, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), Western blotting, nephelometry, nephelometry, immunoradiometry, lateral flow, and immunohistochemistry / cytochemistry.

[0214] However, those skilled in the art will appreciate that several methods can be used to detect or determine the presence or absence of one or more biomarkers, including mass spectrometry approaches such as MS / MS, LC-MS / MS, multiple reaction monitoring (MRM), or SRM, and product ion monitoring (PIM), as well as antibody-based methods such as immunoassays, e.g., Western blot, enzyme-linked immunosorbent assay (ELISA), immunoprecipitation, immunohistochemistry, immunofluorescence, radioimmunoassay, dot blotting, and FACS. Thus, in some embodiments, determining the level of at least one biomarker comprises using an immunoassay and / or mass spectrometry. In additional embodiments, the mass spectrometry method is selected from MS, MS / MS, LC-MS / MS, SRM, PIM, and other such methods known in the art. In other embodiments, the LC-MS / MS further comprises 1D LC-MS / MS, 2D LC-MS / MS, or 3D LC-MS / MS. Immunoassay techniques and protocols are generally known to those skilled in the art (Price and Newman, Principles and Practice of Immunoassay, 2nd ed., Grove's Dictionaries, 1997, and Gosling, Immunoassays: A Practical Approach, Oxford University Press, 2000). A variety of immunoassay techniques can be used, including competitive and non-competitive immunoassays (Self et al., Curr. Opin. Biotechnol., 7:60-65 (1996)).

[0215] In one aspect, the present disclosure relates to a method for determining the probability of preterm birth in a pregnant woman, the method comprising measuring one or more biomarkers in a biological sample from the pregnant woman. In some embodiments, the one or more biomarkers include SHBG. In some embodiments, the one or more biomarkers include IBP4. In some embodiments, the one or more biomarkers include SHBG and IBP4. According to the present disclosure, the method can be used to determine the probability of preterm birth.

[0216] In some embodiments, the methods include immunoassays in which one or more biomarkers are labeled and / or detected by binding of an anti-SHBG or anti-IBP4 antibody described herein. In some embodiments, the detecting anti-SHBG and / or anti-IBP4 antibody can be labeled with different types of labels that can produce different types of signals that can be visualized or detected using a variety of standard procedures, such as luminescence, chemiluminescence, fluorescence, absorbance, radioactivity detection, or by microscopy, imaging, etc. Immunoassays can include immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), Western blotting, nephelometry, nephelometry, immunoradiometry, lateral flow, immunohistochemistry / cytochemistry, and other methods known to those skilled in the art.

[0217] In some embodiments, the method can be used to determine the presence or absence of a biomarker in a sample, and the amount of the biomarker in the sample. The presence or absence or amount of a biomarker in a sample can be determined by comparison with (or as a ratio to) a reference or standard, for example, a sample obtained from a healthy pregnant woman. The amount of a biomarker in a sample can also be determined by comparison with a reference or standard, for example, the amount of an endogenous or recombinant or synthetic biomarker fragment in a reference or control sample. Thus, the amount of a biomarker in a sample need not be quantified in absolute terms, but can be measured in relative terms with respect to a reference or control.

[0218] In another aspect, the present disclosure relates to a method for determining the probability of preeclampsia in a pregnant woman, the method comprising measuring one or more biomarkers in a biological sample from the pregnant woman. In some embodiments, the one or more biomarkers include SHBG. In some embodiments, the one or more biomarkers include IBP4. In some embodiments, the one or more biomarkers include SHBG and IBP4. According to the present disclosure, the method can be used to determine the probability of preeclampsia.

[0219] In another aspect, the present disclosure relates to a method for detecting one or more biomarkers in a biological sample. The method may include obtaining a biological sample, for example, a biological sample (e.g., serum or urine) from a pregnant woman. The method for detecting whether one or more biomarkers are present in the biological sample includes contacting the biological sample containing the one or more biomarkers with an antibody or fragment thereof disclosed herein and detecting binding of the antibody or fragment thereof to the one or more biomarkers. Binding can be detected using a label. Binding can also be detected by mass spectrometry. In some embodiments, the one or more biomarkers include SHBG. In some embodiments, the one or more biomarkers include IBP4. In some embodiments, the one or more biomarkers include SHBG and IBP4.

[0220] In another aspect, the present disclosure provides a method for assessing or predicting the health status of a subject, comprising: (i) obtaining a biological sample containing one or more biomarkers from the subject; (ii) contacting the biological sample with an anti-SHBG antibody or anti-IBP4 antibody, or fragment thereof, described herein, under conditions suitable for the antibody or fragment thereof to bind to the one or more biomarkers; and (iii) detecting binding of the antibody or fragment thereof to the one or more biomarkers, wherein detecting binding of the antibody to the one or more biomarkers indicates the presence of the one or more biomarkers in the biological sample, and not detecting binding of the antibody to the one or more biomarkers indicates the absence of the one or more biomarkers in the biological sample, and the presence or absence of the one or more biomarkers in the biological sample is used to assess or predict the health status of the subject. In some embodiments, the method further comprises quantifying the amount of one or more biomarkers bound to the antibody or fragment thereof in the biological sample, and the amount of the one or more biomarkers is used to assess or predict the health status of the subject. In some embodiments, the method comprises detecting SHBG. In some embodiments, the method comprises detecting IBP4. In some embodiments, the method comprises detecting SHBG and IBP4. In some embodiments, the health condition comprises any one of abnormal glucola, gestational diabetes, hypertension, preeclampsia, intrauterine growth restriction, stillbirth, fetal growth restriction, HELLP syndrome, oligohyramnios, chorioamnionitis, chorioamnionitis, placenta previa, placental acreta, abruption, placental abruption, placental hemorrhage, premature premature rupture of membranes, preterm labor, unfavorable cervix, post-term pregnancy, cholelithiasis, or uterine over distention.

[0221] Thus, in some embodiments, the methods of the present disclosure use an anti-SHBG antibody or fragment thereof comprising a heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 11-22, 115, or 116, and a light chain comprising a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-34. In some embodiments, the heavy chain associates with the light chain to form a binding site for SHBG. In other embodiments, the anti-SHBG antibody or fragment thereof comprises a heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 11, 14, 17, 44, 45, 47, 48, and 50-54, and / or a light chain comprising a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-34. In some embodiments, the heavy chain associates with the light chain to form a binding site for SHBG. In some embodiments, the methods of the present disclosure use an anti-IBP4 antibody or fragment thereof comprising a heavy chain comprising a heavy chain complementarity-determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 75-86, 118, 119, 216-225, 244-258, 284-296, and 312-322, and a light chain comprising a light chain complementarity-determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 87, 24, 89, 90, 91, 93, 94, 95-98, 226-235, 259-264, 272-275, 297-303, and 323-332. In some embodiments, the heavy chain associates with the light chain to form a binding site for IBP4.In other embodiments, the anti-IBP4 antibody or fragment thereof comprises a heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 75-86, 118, 119, 216-225, 244-258, 284-296, and 312-322, and / or a light chain comprising a light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 87, 24, 90, 91, 93, 94, 96, 97, 99, 100, 101, 226-235, 259-264, 272-275, 297-303, and 323-332. In some embodiments, the heavy chain associates with the light chain to form a binding site for IBP4.

[0222] In further embodiments, the present disclosure is described and exemplified with reference to methods for determining the probability of preterm birth or preeclampsia in a pregnant woman, but is equally applicable to methods for predicting abnormal glucose tests, gestational diabetes, hypertension, intrauterine growth retardation, stillbirth, fetal growth restriction, HELLP syndrome, oligohydramnios, chorioamnionitis, chorioamnionitis, placenta previa, placenta accreta, abruption, placental abruption, placental hemorrhage, premature premature rupture of membranes, preterm labor, adverse cervix, post-term pregnancy, cholelithiasis, uterine hyperdistension, and stress.

[0223] In some embodiments, suitable biological samples include, for example, whole blood, plasma, serum, amniotic fluid, vaginal secretions, saliva, and urine. In certain embodiments, the biological sample is serum.

[0224] In some embodiments, the biological sample is obtained at a gestational age of 19-21 weeks. In some embodiments, the pregnant woman is 17-28 weeks pregnant at the time the biological sample is collected, also referred to as GABD (gestational age at blood draw). In other embodiments, the pregnant woman is 16-29 weeks, 17-28 weeks, 18-27 weeks, 19-26 weeks, 20-25 weeks, 21-24 weeks, or 22-23 weeks pregnant at the time the biological sample is collected. In further embodiments, the pregnant woman is about 17-22 weeks, about 16-22 weeks, about 22-25 weeks, about 13-25 weeks, about 26-28 weeks, or ...

Claims

1. An isolated anti-insulin-like growth factor binding protein 4 (IBP4) antibody or fragment thereof, comprising: (i) a heavy chain complementarity determining region 1 (HCDR1) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 75, 78, 81, 84 and 118; and a heavy chain complementarity determining region 2 (HCDR2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 76, 79, 82, 85, and 119; and Heavy chain complementarity determining region 3 (HCDR3) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 77, 80, 83 and 86 and / or a heavy chain comprising (ii) a light chain complementarity-determining region 1 (LCDR1) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 87, 90, 93, and 96; and a light chain complementarity determining region 2 (LCDR2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 24, 91, 94, and 97; and Light chain complementarity determining region 3 (LCDR3) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 89, 95, 98, 99, 100 and 101. light chain containing 1. An isolated anti-insulin-like growth factor binding protein 4 (IBP4) antibody or fragment thereof, comprising:

2. An isolated anti-IBP4 antibody or fragment thereof described in claim 1, wherein the antibody or fragment thereof comprises the heavy chain and the light chain, the heavy chain and the light chain forming an antigen-binding domain for binding to an epitope of IBP4, and the epitope of IBP4 comprises the amino acid sequence of SEQ ID NO:

111. (i) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 75, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 76, and the HCDR3 comprises the amino acid sequence of SEQ ID NO: 77; (ii) the LCDR1 comprises the amino acid sequence of SEQ ID NO: 87, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 24, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 89; or (iii) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 75, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 76, and the HCDR3 comprises the amino acid sequence of SEQ ID NO: 77; (iv) the LCDR1 comprises the amino acid sequence of SEQ ID NO: 87, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 24, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 99; or (v) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 78, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 79, and the HCDR3 comprises the amino acid sequence of SEQ ID NO: 80; (vi) the LCDR1 comprises the amino acid sequence of SEQ ID NO: 90, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 91, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 89; or (vii) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 78, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 79, and the HCDR3 comprises the amino acid sequence of SEQ ID NO: 80; (viii) the LCDR1 comprises the amino acid sequence of SEQ ID NO: 90, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 91, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 99; or (ix) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 81, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 82, and the HCDR3 comprises the amino acid sequence of SEQ ID NO: 83; (x) the LCDR1 comprises the amino acid sequence of SEQ ID NO: 93, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 94, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 95; or (xi) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 81, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 82, and the HCDR3 comprises the amino acid sequence of SEQ ID NO: 83; (xii) the LCDR1 comprises the amino acid sequence of SEQ ID NO: 93, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 94, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 100; or (xiii) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 84, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 85, and the HCDR3 comprises the amino acid sequence of SEQ ID NO: 86; (xiv) the LCDR1 comprises the amino acid sequence of SEQ ID NO: 96, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 97, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 98; or (xv) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 84, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 85, and the HCDR3 comprises the amino acid sequence of SEQ ID NO: 86; (xvi) the LCDR1 comprises the amino acid sequence of SEQ ID NO: 96, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 97, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 101; or (xvii) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 118, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 119, and the HCDR3 comprises the amino acid sequence of SEQ ID NO: 83; (xviii) the LCDR1 comprises the amino acid sequence of SEQ ID NO: 93, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 94, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 89; or (xix) the HCDR1 comprises the amino acid sequence of SEQ ID NO: 118, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 119, and the HCDR3 comprises the amino acid sequence of SEQ ID NO: 83; (xx) the LCDR1 comprises the amino acid sequence of SEQ ID NO: 93, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 94, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 99; 3. The isolated anti-IBP4 antibody or fragment thereof of claim 1 or 2. (i) an HCDR1 comprising the amino acid sequence GYTFTDYY (SEQ ID NO: 75), an HCDR2 comprising the amino acid sequence INPNNGGS (SEQ ID NO: 76), and an HCDR3 comprising the amino acid sequence ARWDGGFDY (SEQ ID NO: 77); and (ii) an LCDR1 comprising the amino acid sequence QNVGTN (SEQ ID NO: 87), an LCDR2 comprising the amino acid sequence SAS (SEQ ID NO: 24), and an LCDR3 comprising the amino acid sequence QQYNX 1 YPLT (SEQ ID NO: 114), wherein X 1 is (i) any naturally occurring amino acid residue, or (ii) isoleucine or serine.

1. An isolated anti-IBP4 antibody or fragment thereof comprising: (i) a heavy chain variable (VH) sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 72 and a light chain variable (VL) sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 74; or (ii) a VH sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the sequence of SEQ ID NO: 107 and a VL sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:

109.

10. The isolated anti-IBP4 antibody or fragment thereof of claim 1, 2, or 4, comprising: (i) a heavy chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 68 or 123, or a light chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 70 or 122; or (ii) a heavy chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 103 or 128, or a light chain comprising an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 105 or 129.

10. The isolated anti-IBP4 antibody or fragment thereof of claim 1, 2, or 4, comprising:

7. An isolated anti-IBP4 antibody or fragment thereof according to any one of claims 1, 2 or 4, wherein the antibody or fragment thereof has a dissociation constant (KD) of about 1x10-7 M or less, 1x10-8 M or less, 1x10-9 M or less, 1x10-10 M or less, 2x10-7 M or less, 2x10-8 M or less, 2x10-9 M or less, 2x10-10 M or less, 5x10-7 M or less, 5x10-8 M or less, 5x10-9 M or less, or 5x10-10 M or less.

8. An isolated anti-IBP4 antibody or fragment thereof described in any one of claims 1, 2 or 4, wherein the antibody or fragment thereof is bound to a solid support.

9. A nucleic acid encoding the heavy chain of an anti-IBP4 antibody or fragment thereof according to claim 1, 2 or 4; and / or A gene encoding the light chain of the anti-IBP4 antibody or fragment thereof of any one of claims 1, 2 or 4. Nucleic acid molecule.

10. A cell containing the nucleic acid molecule described in claim 9.

11. A method for detecting one or more biomarkers in a biological sample, comprising: (i) contacting a biological sample containing one or more biomarkers with an isolated anti-IBP4 antibody or fragment thereof of any one of claims 1, 2 or 4; and (ii) detecting binding of said antibody or said fragment thereof to said one or more biomarkers. A method comprising:

12. A method for detecting the presence of a biomarker as an indicator of a health status of a subject, said method comprising: (i) contacting the isolated anti-IBP4 antibody or fragment thereof of any one of claims 1, 2 or 4 with a biological sample obtained from a subject containing one or more biomarkers under conditions suitable for the antibody or fragment thereof to bind to the one or more biomarkers; and (ii) detecting said binding of said antibody or said fragment thereof to said one or more biomarkers. Including, wherein detecting binding of the antibody to the one or more biomarkers indicates the presence of the one or more biomarkers in the biological sample, and not detecting binding of the antibody to the one or more biomarkers indicates the absence of the one or more biomarkers in the biological sample, and wherein the presence or absence of the one or more biomarkers in the biological sample is used as an indicator of the health status of the subject. method.

13. An anti-IBP4 antibody or fragment thereof according to any one of claims 1, 2, or 4; and Carrier A composition comprising:

14. An anti-IBP4 antibody or fragment thereof according to any one of claims 1, 2 or 4. one or more auxiliary reagents; and Packaging for it wherein the anti-IBP4 antibody or fragment thereof is bound to a solid support.

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