Anticorticotropin-releasing hormone antibodies and their use in congenital adrenal hyperplasia

Abstract

Disclosed herein are antibodies and antigen-binding fragments thereof that specifically bind to corticotropin-releasing hormone (CRH). Also disclosed are methods, pharmaceutical compositions containing such antibodies and antigen-binding fragments, and the use of said anti-corticotropin-releasing hormone (CRH) antibodies and antigen-binding fragments thereof in the treatment of congenital adrenal hyperplasia (CAH) and related disorders.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This PCT application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 234,130, filed August 17, 2021, which is incorporated herein by reference in its entirety.

[0002] Reference to an electronically submitted sequence listing The contents of the electronically submitted Sequence Listing (Name: 4857_001PC01_Seqlisting_ST26; Size: 685,799 bytes; and Creation Date: August 3, 2022) are incorporated herein by reference in their entirety.

[0003] The present disclosure relates to antibodies and antigen-binding fragments thereof that specifically bind to corticotropin-releasing hormone (CRH) and block CRH from binding to its receptor. The present disclosure also relates to pharmaceutical compositions containing such antibodies and antigen-binding fragments thereof, and to the use of anti-CRH antibodies and antigen-binding fragments thereof in the treatment of congenital adrenal hyperplasia and other related disorders and conditions. [Background technology]

[0004] Congenital adrenal hyperplasia (CAH) is a group of rare, inherited, autosomal recessive disorders characterized by a deficiency of one of the enzymes required for hormone production in the adrenal glands. CAH affects the adrenal glands, located at the top of each kidney. Normally, the adrenal glands are responsible for producing three different hormones: (i) corticosteroids, which control the body's response to disease or injury; (ii) mineralocorticoids, which regulate salt and water levels; and (iii) androgens, the male sex hormones. The enzyme deficiency associated with CAH renders the adrenal glands unable to produce one or more of these hormones, which then leads to the overproduction of precursors of other types of hormones in response to this loss.

[0005] The most common cause of CAH is a deficiency of the enzyme 21-hydroxylase. CAH due to 21-hydroxylase deficiency accounts for 95% of all cases and is further divided into two subcategories: (i) classic CAH, which can be subdivided into salt-wasting and simple virilizing forms, and (ii) non-classic CAH. Classic CAH is a more severe form and, if undetected and untreated, can lead to adrenal crisis and death. Non-classic CAH is milder and may or may not exhibit symptoms. 21-hydroxylase deficiency interferes with an individual's ability to produce the hormone cortisol and, in the case of salt-wasting CAH, aldosterone. In addition, the body produces more androgens, which causes various symptoms, such as abnormal reproductive development in female infants. Other, much rarer forms of CAH also exist, including 11-beta-hydroxylase deficiency, 17a-hydroxylase deficiency, 3-beta-hydroxysteroid dehydrogenase deficiency, congenital lipoid adrenal hyperplasia, and p450 oxidoreductase deficiency.

[0006] Classical CAH is a group of autosomal recessive disorders resulting in enzyme deficiency that alters adrenal steroid production due to 21-hydroxylase deficiency, a condition that results in little or no cortisol biosynthesis. One clinical manifestation of cortisol deficiency is a lack of feedback inhibition of pituitary adrenocorticotropic hormone (ACTH) secretion. Increased ACTH levels lead to adrenal hyperplasia, and enzyme mutations cause shunting of cortisol precursor steroids to alternative pathways. Most notably, shunting to androgens leads to virilization and other developmental complications in women, and high ACTH concentrations are associated with the formation of testicular adrenal rest tumors (TARTs) in men. Additionally, because 21-hydroxylase is used in the pathway of mineralocorticoid biosynthesis, many of these patients suffer from aldosterone deficiency, which can lead to dehydration and death due to salt loss. Based on newborn screening, the prevalence of classic 21-hydroxylase deficiency CAH in the general US population has been documented to be 1:10,000 to 1:20,800 (Trakakis et al., Gynecol Endocrinol (2010) 26(1):63-71; Hertzberg et al., Pediatr. (2011) 159(4):555-560).

[0007] Pediatric patients from birth to adolescence, especially females, appear to be the most vulnerable population affected by CAH and represent the subgroup with the greatest unmet medical needs (Cheng and Speiser, Adv. Pediatr. (2012) 59(1):269-281; ​​Merke and Poppas, Lancet Diabetes Endocrinol. (2013) 1(4):341-352). Excessive androgen production in these young patients leads to precocious puberty and adrenocortical onset, altered skeletal maturation patterns, short stature caused by immature epiphyseal plate fusion, and significant hirsutism and acne problems. Although steroid replacement strategies based on physiological dosing of glucocorticoids (e.g., hydrocortisone) and mineralocorticoids (e.g., fludrocortisone) adequately ensure survival, these doses are often inadequate to suppress the accumulation of ACTH and the overproduction of progesterone and androgens (e.g., 17-hydroxyprogesterone [17-OHP], testosterone, free testosterone, androstenedione, 11-oxygenated androgens, or combinations thereof). Uncontrolled symptoms of androgen excess have a substantial impact on the day-to-day functioning and development of these patients.

[0008] Currently, exogenous corticosteroids are the standard of care for treating patients with classic CAH. This treatment is used to correct cortisol deficiency and to reduce excess ACTH levels and androgen excess. However, the dose and duration of steroid use required to suppress ACTH typically far exceeds the normal physiological levels used with cortisol replacement alone. This increased exposure to glucocorticoids can lead to, for example, iatrogenic Cushing's syndrome, increased cardiovascular risk factors, glucose intolerance, reduced growth rate, and decreased bone mineral density in CAH patients (Elnecave et al., J. Pediatr. Endocrinol. Metab. (2008) 21(12):1155-1162; King et al., J. Clin. Endocrinol Metab. (2006) 91(3):865-869; Migeon and Wisniewski, Endocrinol. Metab. Clin. North Am. (2001) 30(1):193-206). Thus, there is currently a need in the art for improved therapies for CAH. Summary of the Invention

[0009] Provided herein is an antibody or antigen-binding fragment thereof that specifically binds to corticotropin-releasing hormone (CRH), the antibody or antigen-binding fragment thereof comprising: (a) a heavy chain variable domain (VH) CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 22 to 35, a VH CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 53 to 78, and a VH CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 113 to 122 or the amino acid sequence of a PDV or GID; and (b) a light chain variable domain (VL) CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 152 to 174, a VL CDR2 comprising the amino acid sequence of any one of amino acid sequences 193 to 198, and a VL CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 223 to 232.

[0010] In one embodiment, the antibody comprises: (a) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 53, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 152, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223; (b) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 54, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 152, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (c) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 55, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 152, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (d) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 53, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 153, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (e) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 54, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 153, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (f) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 55, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 153, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223.(g) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 54, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 154, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (h) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 54, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 155, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (i) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 54, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 156, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (j) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 54, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 157, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (k) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 55, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 154, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (l) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 55, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 155, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223.(m) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 55, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 156, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (n) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 55, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 157, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (o) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 56, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 154, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (p) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 56, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 155, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (q) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 56, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 156, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223. (r) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 56, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 157, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223.(s) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 23, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 57, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 114, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 158, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 224. (t) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 23, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 58, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 114, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 158, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 224. (u) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 23, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 59, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 114, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 158, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 224. (v) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 23, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 60, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 114, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 158, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 224. (w) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 24, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 61, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 115, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 159, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 194, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 225. (x) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 25, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 62, VH CDR3 comprises the amino acid sequence of PDV, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 160, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 195, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 226.(y) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 24, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 63, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 115, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 161, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 196, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 225. (z) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 26, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 64, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 116, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 162, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 227. (aa) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 24, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 63, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 115, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 163, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 196, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 225. (bb) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 24, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 65, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 115, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 161, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 196, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 225. (cc) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 27, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 66, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 117, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 164, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 228. (dd) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 28, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 67, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 118, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 165, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 227. (ee) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 68, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 119, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 166, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 227. (ff) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 30, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 69, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 120, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 167, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 229.(gg) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 70, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 166, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (hh) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 31, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 71, VH CDR3 comprises the amino acid sequence of GID, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 168, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 198, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 231. (ii) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 32, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 72, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 118, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 166, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 227. (jj) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 33, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 73, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 118, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 169, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 227. (kk) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 34, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 71, VH CDR3 comprises the amino acid sequence of GID, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 168, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 198, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 231. (ll) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 30, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 74, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 122, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 170, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 232.(mm) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 35, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 71, and VH CDR3 comprises the amino acid sequence of GID, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 168, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 198, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 231. (nn) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 75, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 171, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (oo) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 76, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 171, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (pp) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 77, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 171, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (qq) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 75, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 172, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (rr) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 75, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 173, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230.(ss) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 76, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 172, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (tt) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 77, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 173, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (uu) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 76, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 173, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (vv) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 77, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 172, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (ww) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 78, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 172, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (xx) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 78, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 173, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230.(yy) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 76, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 174, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (zz) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 77, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 174, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (aaa) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 78, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 174, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (bbb) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 78, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 171, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230. (ccc) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 29, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 75, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 121, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 174, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 230; or (ddd) VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, VH CDR2 comprises the amino acid sequence of SEQ ID NO: 53, VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, VL CDR1 comprises the amino acid sequence of SEQ ID NO: 152, VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223.

[0011] In another aspect, the present invention relates to an antibody or antigen-binding fragment thereof comprising: (a) a VH comprising an amino acid sequence at least 90%, 95%, 99%, or 100% identical to any one of SEQ ID NOs: 240-295, 459, and 461; and / or (b) a VL comprising an amino acid sequence at least 90%, 95%, 99%, or 100% identical to any one of SEQ ID NOs: 296-347 and 462. In one embodiment, the antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 99%, or 100% identical to: (a) the VH amino acid sequence of SEQ ID NO: 240 and / or the VL amino acid sequence of SEQ ID NO: 296, (b) the VH amino acid sequence of SEQ ID NO: 241 and / or the VL amino acid sequence of SEQ ID NO: 296, (c) the VH amino acid sequence of SEQ ID NO: 242 and / or the VL amino acid sequence of SEQ ID NO: 296, (d) the VH amino acid sequence of SEQ ID NO: 240 and / or the VL amino acid sequence of SEQ ID NO: 297, (e) the VH amino acid sequence of SEQ ID NO: 241 and / or the VL amino acid sequence of SEQ ID NO: 297, (f) the VH amino acid sequence of SEQ ID NO: 242 and / or the VL amino acid sequence of SEQ ID NO: 297, (g) the VH amino acid sequence of SEQ ID NO: 241 and / or the VL amino acid sequence of SEQ ID NO: 298, (h) the VH amino acid sequence of SEQ ID NO: 241. (i) the VH amino acid sequence of SEQ ID NO: 241 and the VL amino acid sequence of SEQ ID NO: 300, (j) the VH amino acid sequence of SEQ ID NO: 241 and / or the VL amino acid sequence of SEQ ID NO: 301, (k) the VH amino acid sequence of SEQ ID NO: 242 and / or the VL amino acid sequence of SEQ ID NO: 298, (l) the VH amino acid sequence of SEQ ID NO: 242 and / or the VL amino acid sequence of SEQ ID NO: 299, (m) the VH amino acid sequence of SEQ ID NO: 242 and / or the VL amino acid sequence of SEQ ID NO: 300, (n) the VH amino acid sequence of SEQ ID NO: 242 and / or the VL amino acid sequence of SEQ ID NO: 301, (o) the VH amino acid sequence of SEQ ID NO: 243 and / or the VL amino acid sequence of SEQ ID NO: 298, (p) the VH amino acid sequence of SEQ ID NO: 243 and / or the VL amino acid sequence of SEQ ID NO: 299, (q) the VH amino acid sequence of SEQ ID NO: 243,and / or the VL amino acid sequence of SEQ ID NO: 300, (r) the VH amino acid sequence of SEQ ID NO: 243, and / or the VL amino acid sequence of SEQ ID NO: 301, (s) the VH amino acid sequence of SEQ ID NO: 244, and / or the VL amino acid sequence of SEQ ID NO: 302, (t) the VH amino acid sequence of SEQ ID NO: 245, and / or the VL amino acid sequence of SEQ ID NO: 302, (u) the VH amino acid sequence of SEQ ID NO: 246, and / or the VL amino acid sequence of SEQ ID NO: 302, (v) SEQ ID NO: 247 and / or the VL amino acid sequence of SEQ ID NO: 302, (w) the VH amino acid sequence of SEQ ID NO: 248 and / or the VL amino acid sequence of SEQ ID NO: 303, (x) the VH amino acid sequence of SEQ ID NO: 249 and / or the VL amino acid sequence of SEQ ID NO: 304, (y) the VH amino acid sequence of SEQ ID NO: 250 and / or the VL amino acid sequence of SEQ ID NO: 305, (z) the VH amino acid sequence of SEQ ID NO: 251 and / or the VL amino acid sequence of SEQ ID NO: 306, (aa) the VH amino acid sequence of SEQ ID NO: 252 and / or the VL amino acid sequence of SEQ ID NO: 305, (bb) the VH amino acid sequence of SEQ ID NO: 252 and / or the VL amino acid sequence of SEQ ID NO: 307, (cc) the VH amino acid sequence of SEQ ID NO: 253 and / or the VL amino acid sequence of SEQ ID NO: 308, (dd) the VH amino acid sequence of SEQ ID NO: 254 and / or the VL amino acid sequence of SEQ ID NO: 309, (ee) the VH amino acid sequence of SEQ ID NO: 255 and / or the VL amino acid sequence of SEQ ID NO: 310, (ff) the VH amino acid sequence of SEQ ID NO: 255, and / or the VL amino acid sequence of SEQ ID NO: 311, (gg) the VH amino acid sequence of SEQ ID NO: 255, and / or the VL amino acid sequence of SEQ ID NO: 312, (hh) the VH amino acid sequence of SEQ ID NO: 256, and / or the VL amino acid sequence of SEQ ID NO: 310, (ii) the VH amino acid sequence of SEQ ID NO: 256, and / or the VL amino acid sequence of SEQ ID NO: 311, (jj) the VH amino acid sequence of SEQ ID NO: 256, and / or the VL amino acid sequence of SEQ ID NO: 312, (kk) the VH amino acid sequence of SEQ ID NO: 257, and / or the VL amino acid sequence of SEQ ID NO: 310 sequence, (ll) the VH amino acid sequence of SEQ ID NO: 257 and / or the VL amino acid sequence of SEQ ID NO: 311, (mm) the VH amino acid sequence of SEQ ID NO: 257 and / or the VL amino acid sequence of SEQ ID NO: 312, (nn) the VH amino acid sequence of SEQ ID NO: 258 and / or the VL amino acid sequence of SEQ ID NO: 310, (oo) the VH amino acid sequence of SEQ ID NO: 258 and / or the VL amino acid sequence of SEQ ID NO: 311, (pp) the VH amino acid sequence of SEQ ID NO: 258 and / or the VL amino acid sequence of SEQ ID NO: 312, (qq) the VH amino acid sequence of SEQ ID NO: 2 , and / or the VL amino acid sequence of SEQ ID NO: 310, (rr) the VH amino acid sequence of SEQ ID NO: 2, and / or the VL amino acid sequence of SEQ ID NO: 311, (ss) the VH amino acid sequence of SEQ ID NO: 2, and / or the VL amino acid sequence of SEQ ID NO: 312, (tt) the VH amino acid sequence of SEQ ID NO: 260, and / or the VL amino acid sequence of SEQ ID NO: 317, (uu) the VH amino acid sequence of SEQ ID NO: 260, and / or the VL amino acid sequence of SEQ ID NO: 313, (vv) the VH amino acid sequence of SEQ ID NO: 261, and / or the VL amino acid sequence of SEQ ID NO: 314,(ww) the VH amino acid sequence of SEQ ID NO: 261 and / or the VL amino acid sequence of SEQ ID NO: 315, (xx) the VH amino acid sequence of SEQ ID NO: 261 and / or the VL amino acid sequence of SEQ ID NO: 316, (yy) the VH amino acid sequence of SEQ ID NO: 262 and / or the VL amino acid sequence of SEQ ID NO: 317, (zz) the VH amino acid sequence of SEQ ID NO: 261 and / or the VL amino acid sequence of SEQ ID NO: 317, (aaa) the VH amino acid sequence of SEQ ID NO: 263 and / or the VL amino acid sequence of SEQ ID NO: 317, (bbb) the VH amino acid sequence of SEQ ID NO: 264 and / or the VL amino acid sequence of SEQ ID NO: 317, (ccc) the VH amino acid sequence of SEQ ID NO: 262 and / or the VL amino acid sequence of SEQ ID NO: 318, (ddd) the VH amino acid sequence of SEQ ID NO: 263 and / or the VL amino acid sequence of SEQ ID NO: 318, (eee) the VH amino acid sequence of SEQ ID NO: 264 and / or the VL amino acid sequence of SEQ ID NO: 318, (fff) the VH amino acid sequence of SEQ ID NO: 260 (iii) the VH amino acid sequence of SEQ ID NO: 260 and / or the VL amino acid sequence of SEQ ID NO: 319, (jjj) the VH amino acid sequence of SEQ ID NO: 260 and / or the VL amino acid sequence of SEQ ID NO: 314, (kkk) the VH amino acid sequence of SEQ ID NO: 260 and / or the VL amino acid sequence of SEQ ID NO: 3 15 VL amino acid sequence, (lll) the VH amino acid sequence of SEQ ID NO: 260 and / or the VL amino acid sequence of SEQ ID NO: 316, (mmm) the VH amino acid sequence of SEQ ID NO: 261 and / or the VL amino acid sequence of SEQ ID NO: 313, (nnn) the VH amino acid sequence of SEQ ID NO: 261 and / or the VL amino acid sequence of SEQ ID NO: 318, (ooo) the VH amino acid sequence of SEQ ID NO: 261 and / or the VL amino acid sequence of SEQ ID NO: 319, (ppp) the VH amino acid sequence of SEQ ID NO: 265 and / or the VL amino acid sequence of SEQ ID NO: 320, (qqq) the VH amino acid sequence of SEQ ID NO: 266 and / or the VL amino acid sequence of SEQ ID NO: 321, (rrr) the VH amino acid sequence of SEQ ID NO: 267 and / or the VL amino acid sequence of SEQ ID NO: 322, (sss) the VH amino acid sequence of SEQ ID NO: 268 and / or the VL amino acid sequence of SEQ ID NO: 323, (ttt) the VH amino acid sequence of SEQ ID NO: 269 and / or the VL amino acid sequence of SEQ ID NO: 324, (uuu) the VH amino acid sequence of SEQ ID NO: 270 and / or the VL amino acid sequence of SEQ ID NO: 325, (vvv) the VH amino acid sequence of SEQ ID NO: 271,and / or the VL amino acid sequence of SEQ ID NO: 326, (www) the VH amino acid sequence of SEQ ID NO: 272, and / or the VL amino acid sequence of SEQ ID NO: 331, (xxx) the VH amino acid sequence of SEQ ID NO: 272, and / or the VL amino acid sequence of SEQ ID NO: 327, (yyy) the VH amino acid sequence of SEQ ID NO: 273, and / or the VL amino acid sequence of SEQ ID NO: 328, (zzz) the VH amino acid sequence of SEQ ID NO: 273, and / or the VL amino acid sequence of SEQ ID NO: 329, (aaaa) the VH amino acid sequence of SEQ ID NO: 273 and / or the VL amino acid sequence of SEQ ID NO: 330, (bbbb) the VH amino acid sequence of SEQ ID NO: 273 and / or the VL amino acid sequence of SEQ ID NO: 331, (cccc) the VH amino acid sequence of SEQ ID NO: 274 and / or the VL amino acid sequence of SEQ ID NO: 331, (dddd) the VH amino acid sequence of SEQ ID NO: 275 and / or the VL amino acid sequence of SEQ ID NO: 331, (eeee) the VH amino acid sequence of SEQ ID NO: 276 and / or the VL amino acid sequence of SEQ ID NO: 331, (ffff) SEQ ID NO: 2 77 and / or the VL amino acid sequence of SEQ ID NO: 331, (gggg) the VH amino acid sequence of SEQ ID NO: 272 and / or the VL amino acid sequence of SEQ ID NO: 332, (hhhh) the VH amino acid sequence of SEQ ID NO: 276 and / or the VL amino acid sequence of SEQ ID NO: 332, (iiii) the VH amino acid sequence of SEQ ID NO: 275 and / or the VL amino acid sequence of SEQ ID NO: 328, (jjjj) the VH amino acid sequence of SEQ ID NO: 275 and / or the VL amino acid sequence of SEQ ID NO: 330, (kkkk) the VH amino acid sequence of SEQ ID NO: 277, and / or the VL amino acid sequence of SEQ ID NO: 328, (llll) the VH amino acid sequence of SEQ ID NO: 277, and / or the VL amino acid sequence of SEQ ID NO: 330, (mmmm) the VH amino acid sequence of SEQ ID NO: 272, and / or the VL amino acid sequence of SEQ ID NO: 333, (nnnn) the VH amino acid sequence of SEQ ID NO: 272, and / or the VL amino acid sequence of SEQ ID NO: 328, (oooo) the VH amino acid sequence of SEQ ID NO: 272, and / or the VL amino acid sequence of SEQ ID NO: 329, (pppp) the VH amino acid sequence of SEQ ID NO: 272, and / or SEQ ID NO: 3 30, (qqqq) the VH amino acid sequence of SEQ ID NO: 273, and / or the VL amino acid sequence of SEQ ID NO: 327, (rrrr) the VH amino acid sequence of SEQ ID NO: 273, and / or the VL amino acid sequence of SEQ ID NO: 332, (ssss) the VH amino acid sequence of SEQ ID NO: 273, and / or the VL amino acid sequence of SEQ ID NO: 333, (tttt) the VH amino acid sequence of SEQ ID NO: 278, and / or the VL amino acid sequence of SEQ ID NO: 334, (uuuu) the VH amino acid sequence of SEQ ID NO: 279, and / or the VL amino acid sequence of SEQ ID NO: 323,(vvvv) the VH amino acid sequence of SEQ ID NO: 280 and / or the VL amino acid sequence of SEQ ID NO: 339, (wwww) the VH amino acid sequence of SEQ ID NO: 280 and / or the VL amino acid sequence of SEQ ID NO: 335, (xxxx) the VH amino acid sequence of SEQ ID NO: 281 and / or the VL amino acid sequence of SEQ ID NO: 336, (yyyy) the VH amino acid sequence of SEQ ID NO: 281 and / or the VL amino acid sequence of SEQ ID NO: 337, (zzzz) the VH amino acid sequence of SEQ ID NO: 281 and / or the VL amino acid sequence of SEQ ID NO: 338, (aaaaa) the VH amino acid sequence of SEQ ID NO: 282 and / or the VL amino acid sequence of SEQ ID NO: 339, (bbbbb) the VH amino acid sequence of SEQ ID NO: 281 and / or the VL amino acid sequence of SEQ ID NO: 339, (ccccc) the VH amino acid sequence of SEQ ID NO: 283 and / or the VL amino acid sequence of SEQ ID NO: 339, (ddddd) the VH amino acid sequence of SEQ ID NO: 284 and / or the VL amino acid sequence of SEQ ID NO: 339, (eeeee) the VH amino acid sequence of SEQ ID NO: 285 and / or the VL amino acid sequence of SEQ ID NO: 339, (fffff ) the VH amino acid sequence of SEQ ID NO: 286 and / or the VL amino acid sequence of SEQ ID NO: 339, (ggggg) the VH amino acid sequence of SEQ ID NO: 282 and / or the VL amino acid sequence of SEQ ID NO: 340, (hhhhh) the VH amino acid sequence of SEQ ID NO: 280 and / or the VL amino acid sequence of SEQ ID NO: 340, (iii) the VH amino acid sequence of SEQ ID NO: 283 and / or the VL amino acid sequence of SEQ ID NO: 340, (jjjjj) the VH amino acid sequence of SEQ ID NO: 284 and / or the VL amino acid sequence of SEQ ID NO: 340, (kkkkk) SEQ ID NO: 2 85 and / or the VL amino acid sequence of SEQ ID NO: 340, (lllll) the VH amino acid sequence of SEQ ID NO: 286 and / or the VL amino acid sequence of SEQ ID NO: 340, (mmmmm) the VH amino acid sequence of SEQ ID NO: 284 and / or the VL amino acid sequence of SEQ ID NO: 336, (nnnnn) the VH amino acid sequence of SEQ ID NO: 284 and / or the VL amino acid sequence of SEQ ID NO: 338, (ooooo) the VH amino acid sequence of SEQ ID NO: 286 and / or the VL amino acid sequence of SEQ ID NO: 336, (ppppp) the VH amino acid sequence of SEQ ID NO: 286 the amino acid sequence of SEQ ID NO: 286 and / or the VL amino acid sequence of SEQ ID NO: 342, (uuuuu) the VH amino acid sequence of SEQ ID NO: 286,and / or the VL amino acid sequence of SEQ ID NO: 343, (vvvvv) the VH amino acid sequence of SEQ ID NO: 287, and / or the VL amino acid sequence of SEQ ID NO: 342, (ww) the VH amino acid sequence of SEQ ID NO: 288, and / or the VL amino acid sequence of SEQ ID NO: 343, (xxxxx) the VH amino acid sequence of SEQ ID NO: 287, and / or the VL amino acid sequence of SEQ ID NO: 343, (yyyyy) the VH amino acid sequence of SEQ ID NO: 288, and / or the VL amino acid sequence of SEQ ID NO: 342, (zzzzz) the VH amino acid sequence of SEQ ID NO: 289, and / or the VL amino acid sequence of SEQ ID NO: 342, (aaaaaa) the VH amino acid sequence of SEQ ID NO: 289 and / or the VL amino acid sequence of SEQ ID NO: 343, (bbbbbb) the VH amino acid sequence of SEQ ID NO: 287 and / or the VL amino acid sequence of SEQ ID NO: 344, (cccccc) the VH amino acid sequence of SEQ ID NO: 288 and / or the VL amino acid sequence of SEQ ID NO: 344, (dddddd) the VH amino acid sequence of SEQ ID NO: 280 and / or the VL amino acid sequence of SEQ ID NO: 336, (eeeeee) the VH amino acid sequence of SEQ ID NO: 289 and / or the VL amino acid sequence of SEQ ID NO: 344, (ffffff) the VH amino acid sequence of SEQ ID NO: 289 and / or the VL amino acid sequence of SEQ ID NO: 340, (gggggg) the VH amino acid sequence of SEQ ID NO: 286 and / or the VL amino acid sequence of SEQ ID NO: 344, (hhhhhh) the VH amino acid sequence of SEQ ID NO: 290 and / or the VL amino acid sequence of SEQ ID NO: 336, (iiiiii) the VH amino acid sequence of SEQ ID NO: 291 and / or the VL amino acid sequence of SEQ ID NO: 336, (jjjjjj) the VH amino acid sequence of SEQ ID NO: 292 and / or the VL amino acid sequence of SEQ ID NO: 336, (kkkkkk) the VH amino acid sequence of SEQ ID NO: 290 and / or the VL amino acid sequence of SEQ ID NO: 345, (llllll) the VH amino acid sequence of SEQ ID NO: 291 and / or the VL amino acid sequence of SEQ ID NO: 345, (mmmmmm) the VH amino acid sequence of SEQ ID NO: 292 and / or the VL amino acid sequence of SEQ ID NO: 345, (nnnnnn) the VH amino acid sequence of SEQ ID NO: 290 and / or the VL amino acid sequence of SEQ ID NO: 346, (oooooo) the VH amino acid sequence of SEQ ID NO: 280 and / or the VL amino acid sequence of SEQ ID NO: 337, (pppppp) the VH amino acid sequence of SEQ ID NO: 291 and / or the VL amino acid sequence of SEQ ID NO: 346, (qqqqqq) the VH amino acid sequence of SEQ ID NO: 292 and / or the VL amino acid sequence of SEQ ID NO: 346, (rrrrrr) the VH amino acid sequence of SEQ ID NO: 290 and / or the VL amino acid sequence of SEQ ID NO: 347, (ssssss) the VH amino acid sequence of SEQ ID NO: 291 and / or the VL amino acid sequence of SEQ ID NO: 347, (tttttt) the VH amino acid sequence of SEQ ID NO: 292 and / or the VL amino acid sequence of SEQ ID NO: 347,(uuuuuu) the VH amino acid sequence of SEQ ID NO: 293 and / or the VL amino acid sequence of SEQ ID NO: 340, (vvvvvv) the VH amino acid sequence of SEQ ID NO: 294 and / or the VL amino acid sequence of SEQ ID NO: 340, (wwwwww) the VH amino acid sequence of SEQ ID NO: 295 and / or the VL amino acid sequence of SEQ ID NO: 340, (xxxxxx) the VH amino acid sequence of SEQ ID NO: 293 and / or the VL amino acid sequence of SEQ ID NO: 343, (yyyyyy) the VH amino acid sequence of SEQ ID NO: 294 and / or the VL amino acid sequence of SEQ ID NO: 343, (zzzzzz) the VH amino acid sequence of SEQ ID NO: 280 and / or the VL amino acid sequence of SEQ ID NO: 338, (aaaaaaa) the VH amino acid sequence of SEQ ID NO: 295 and / or the VL amino acid sequence of SEQ ID NO: 343, (bbbbbbb) the VH amino acid sequence of SEQ ID NO: 293 and / or the VL amino acid sequence of SEQ ID NO: 342, (ccccccc) the VH amino acid sequence of SEQ ID NO: 294 and / or the VL amino acid sequence of SEQ ID NO: 342, (ddddddd) the VH amino acid sequence of SEQ ID NO: 295 and / or the VL amino acid sequence of SEQ ID NO: 342, (eeeeeee) the VH amino acid sequence of SEQ ID NO: 293, and / or the VL amino acid sequence of SEQ ID NO: 344, (fffffff) the VH amino acid sequence of SEQ ID NO: 294, and / or the VL amino acid sequence of SEQ ID NO: 344, (ggggggg) the VH amino acid sequence of SEQ ID NO: 295, and / or the VL amino acid sequence of SEQ ID NO: 344, (hhhhhhh) the VH amino acid sequence of SEQ ID NO: 281, and / or the VL amino acid sequence of SEQ ID NO: 345, (iiiiiii) the VH amino acid sequence of SEQ ID NO: 281, and / or the VL amino acid sequence of SEQ ID NO: 346, (j jjjjjj) the VH amino acid sequence of SEQ ID NO: 281, and / or the VL amino acid sequence of SEQ ID NO: 347, (kkkkkkk) the VH amino acid sequence of SEQ ID NO: 281, and / or the VL amino acid sequence of SEQ ID NO: 335, (lllllll) the VH amino acid sequence of SEQ ID NO: 283, and / or the VL amino acid sequence of SEQ ID NO: 343, (mmmmmmm) the VH amino acid sequence of SEQ ID NO: 283, and / or the VL amino acid sequence of SEQ ID NO: 342, (nnnnnnn) the VH amino acid sequence of SEQ ID NO: 283, and and / or the VL amino acid sequence of SEQ ID NO: 344, (ooooooo) the VH amino acid sequence of SEQ ID NO: 281, and / or the VL amino acid sequence of SEQ ID NO: 340, (ppppppp) the VH amino acid sequence of SEQ ID NO: 281, and / or the VL amino acid sequence of SEQ ID NO: 341, (qqqqqqq) the VH amino acid sequence of SEQ ID NO: 461, and / or the VL amino acid sequence of SEQ ID NO: 462, or (rrrrrrr) the VH amino acid sequence of SEQ ID NO: 459, and / or the VL amino acid sequence of SEQ ID NO: 296.

[0012] In another aspect, the present invention relates to an antibody or antigen-binding fragment thereof comprising: (a) a heavy chain comprising an amino acid sequence at least 90%, 95%, 99%, or 100% identical to any one of SEQ ID NOs: 348-403, 460, and 463; and / or (b) a light chain comprising an amino acid sequence at least 90%, 95%, 99%, or 100% identical to any one of SEQ ID NOs: 404-455 and 464. In one embodiment, the antibody or antigen-binding fragment thereof comprises an amino acid sequence that is at least 90%, 95%, 99%, or 100% identical to: (a) the heavy chain amino acid sequence of SEQ ID NO: 348 and / or the light chain amino acid sequence of SEQ ID NO: 404, (b) the heavy chain amino acid sequence of SEQ ID NO: 349 and / or the light chain amino acid sequence of SEQ ID NO: 404, (c) the heavy chain amino acid sequence of SEQ ID NO: 350 and / or the light chain amino acid sequence of SEQ ID NO: 404, (d) the heavy chain amino acid sequence of SEQ ID NO: 348 and / or the light chain amino acid sequence of SEQ ID NO: 405, (e) the heavy chain amino acid sequence of SEQ ID NO: 349 and / or the light chain amino acid sequence of SEQ ID NO: 405, (f) the heavy chain amino acid sequence of SEQ ID NO: 350 and / or the light chain amino acid sequence of SEQ ID NO: 405, (g) the heavy chain amino acid sequence of SEQ ID NO: 349 and / or the light chain amino acid sequence of SEQ ID NO: 406, (h) the heavy chain amino acid sequence of SEQ ID NO: 349. (i) the heavy chain amino acid sequence of SEQ ID NO: 349 and / or the light chain amino acid sequence of SEQ ID NO: 408, (j) the heavy chain amino acid sequence of SEQ ID NO: 349 and / or the light chain amino acid sequence of SEQ ID NO: 409, (k) the heavy chain amino acid sequence of SEQ ID NO: 350 and / or the light chain amino acid sequence of SEQ ID NO: 406, (l) the heavy chain amino acid sequence of SEQ ID NO: 350 and / or the light chain amino acid sequence of SEQ ID NO: 407, (m) the heavy chain amino acid sequence of SEQ ID NO: 350 and / or the light chain amino acid sequence of SEQ ID NO: 408, (n) the heavy chain amino acid sequence of SEQ ID NO: 350 and / or the light chain amino acid sequence of SEQ ID NO: 409, (o) the heavy chain amino acid sequence of SEQ ID NO: 351 and / or the light chain amino acid sequence of SEQ ID NO: 406, (p) the heavy chain amino acid sequence of SEQ ID NO: 351 and / or the light chain amino acid sequence of SEQ ID NO: 407, (q) the heavy chain amino acid sequence of SEQ ID NO: 351,and / or the light chain amino acid sequence of SEQ ID NO: 408, (r) the heavy chain amino acid sequence of SEQ ID NO: 351, and / or the light chain amino acid sequence of SEQ ID NO: 409, (s) the heavy chain amino acid sequence of SEQ ID NO: 352, and / or the light chain amino acid sequence of SEQ ID NO: 410, (t) the heavy chain amino acid sequence of SEQ ID NO: 353, and / or the light chain amino acid sequence of SEQ ID NO: 410, (u) the heavy chain amino acid sequence of SEQ ID NO: 354, and / or the light chain amino acid sequence of SEQ ID NO: 410, (v) SEQ ID NO: 355 and / or the light chain amino acid sequence of SEQ ID NO: 410, (w) the heavy chain amino acid sequence of SEQ ID NO: 356 and / or the light chain amino acid sequence of SEQ ID NO: 411, (x) the heavy chain amino acid sequence of SEQ ID NO: 357 and / or the light chain amino acid sequence of SEQ ID NO: 412, (y) the heavy chain amino acid sequence of SEQ ID NO: 358 and / or the light chain amino acid sequence of SEQ ID NO: 413, (z) the heavy chain amino acid sequence of SEQ ID NO: 359 and / or the light chain amino acid sequence of SEQ ID NO: 414, (aa) the heavy chain amino acid sequence of SEQ ID NO: 360 and / or the light chain amino acid sequence of SEQ ID NO: 413, (bb) the heavy chain amino acid sequence of SEQ ID NO: 360 and / or the light chain amino acid sequence of SEQ ID NO: 415, (cc) the heavy chain amino acid sequence of SEQ ID NO: 361 and / or the light chain amino acid sequence of SEQ ID NO: 416, (dd) the heavy chain amino acid sequence of SEQ ID NO: 362 and / or the light chain amino acid sequence of SEQ ID NO: 417, (ee) the heavy chain amino acid sequence of SEQ ID NO: 363 and / or the light chain amino acid sequence of SEQ ID NO: 418, (ff) the heavy chain amino acid sequence of SEQ ID NO: 363, and (gg) the heavy chain amino acid sequence of SEQ ID NO: 363 and / or the light chain amino acid sequence of SEQ ID NO: 420, (hh) the heavy chain amino acid sequence of SEQ ID NO: 364 and / or the light chain amino acid sequence of SEQ ID NO: 418, (ii) the heavy chain amino acid sequence of SEQ ID NO: 364 and / or the light chain amino acid sequence of SEQ ID NO: 419, (jj) the heavy chain amino acid sequence of SEQ ID NO: 364 and / or the light chain amino acid sequence of SEQ ID NO: 420, (kk) the heavy chain amino acid sequence of SEQ ID NO: 365 and / or the light chain amino acid sequence of SEQ ID NO: 418, (ll) the heavy chain amino acid sequence of SEQ ID NO: 365 and / or the light chain amino acid sequence of SEQ ID NO: 419, (mm) the heavy chain amino acid sequence of SEQ ID NO: 365 and / or the light chain amino acid sequence of SEQ ID NO: 420, (nn) the heavy chain amino acid sequence of SEQ ID NO: 366 and / or the light chain amino acid sequence of SEQ ID NO: 418, (oo) the heavy chain amino acid sequence of SEQ ID NO: 366 and / or the light chain amino acid sequence of SEQ ID NO: 419, (pp) the heavy chain amino acid sequence of SEQ ID NO: 366 and / or the light chain amino acid sequence of SEQ ID NO: 420, (qq) the heavy chain amino acid sequence of SEQ ID NO: 367, and (rr) the heavy chain amino acid sequence of SEQ ID NO: 367 and / or the light chain amino acid sequence of SEQ ID NO: 419, (ss) the heavy chain amino acid sequence of SEQ ID NO: 367 and / or the light chain amino acid sequence of SEQ ID NO: 420, (tt) the heavy chain amino acid sequence of SEQ ID NO: 368 and / or the light chain amino acid sequence of SEQ ID NO: 425, (uu) the heavy chain amino acid sequence of SEQ ID NO: 368 and / or the light chain amino acid sequence of SEQ ID NO: 421, (vv) the heavy chain amino acid sequence of SEQ ID NO: 369 and / or the light chain amino acid sequence of SEQ ID NO: 422,(ww) the heavy chain amino acid sequence of SEQ ID NO: 369 and / or the light chain amino acid sequence of SEQ ID NO: 423, (xx) the heavy chain amino acid sequence of SEQ ID NO: 369 and / or the light chain amino acid sequence of SEQ ID NO: 424, (yy) the heavy chain amino acid sequence of SEQ ID NO: 370 and / or the light chain amino acid sequence of SEQ ID NO: 425, (zz) the heavy chain amino acid sequence of SEQ ID NO: 369 and / or the light chain amino acid sequence of SEQ ID NO: 425, (aaa) the heavy chain amino acid sequence of SEQ ID NO: 371 and / or the light chain amino acid sequence of SEQ ID NO: 425, (bbb) the heavy chain amino acid sequence of SEQ ID NO: 372 and / or the light chain amino acid sequence of SEQ ID NO: 425, (ccc) the heavy chain amino acid sequence of SEQ ID NO: 370 and / or the light chain amino acid sequence of SEQ ID NO: 426, (ddd) the heavy chain amino acid sequence of SEQ ID NO: 371 and / or the light chain amino acid sequence of SEQ ID NO: 426, (eee) the heavy chain amino acid sequence of SEQ ID NO: 372 and / or the light chain amino acid sequence of SEQ ID NO: 426, (fff) the heavy chain amino acid sequence of SEQ ID NO: 368 (iii) the heavy chain amino acid sequence of SEQ ID NO: 368 and / or the light chain amino acid sequence of SEQ ID NO: 427; (jjj) the heavy chain amino acid sequence of SEQ ID NO: 368 and / or the light chain amino acid sequence of SEQ ID NO: 422; (kkk) the heavy chain amino acid sequence of SEQ ID NO: 368 and / or the light chain amino acid sequence of SEQ ID NO: 426; (ggg) the heavy chain amino acid sequence of SEQ ID NO: 372 and / or the light chain amino acid sequence of SEQ ID NO: 422; (hhh) the heavy chain amino acid sequence of SEQ ID NO: 372 and / or the light chain amino acid sequence of SEQ ID NO: 424; (iii) the heavy chain amino acid sequence of SEQ ID NO: 368 and / or the light chain amino acid sequence of SEQ ID NO: 427; (jjj) the heavy chain amino acid sequence of SEQ ID NO: 368 and / or the light chain amino acid sequence of SEQ ID NO: 422; (kkk) the heavy chain amino acid sequence of SEQ ID NO: 368 and / or the light chain amino acid sequence of SEQ ID NO: 426; 23, (lll) the heavy chain amino acid sequence of SEQ ID NO: 368 and / or the light chain amino acid sequence of SEQ ID NO: 424, (mmm) the heavy chain amino acid sequence of SEQ ID NO: 369 and / or the light chain amino acid sequence of SEQ ID NO: 421, (nnn) the heavy chain amino acid sequence of SEQ ID NO: 369 and / or the light chain amino acid sequence of SEQ ID NO: 426, (ooo) the heavy chain amino acid sequence of SEQ ID NO: 369 and / or the light chain amino acid sequence of SEQ ID NO: 427, (ppp) the heavy chain amino acid sequence of SEQ ID NO: 373 and / or the light chain amino acid sequence of SEQ ID NO: 428, (qqq) a heavy chain amino acid sequence of SEQ ID NO: 374 and / or a light chain amino acid sequence of SEQ ID NO: 429, (rrr) a heavy chain amino acid sequence of SEQ ID NO: 375 and / or a light chain amino acid sequence of SEQ ID NO: 430, (sss) a heavy chain amino acid sequence of SEQ ID NO: 376 and / or a light chain amino acid sequence of SEQ ID NO: 431, (ttt) a heavy chain amino acid sequence of SEQ ID NO: 377 and / or a light chain amino acid sequence of SEQ ID NO: 432, (uuu) a heavy chain amino acid sequence of SEQ ID NO: 378 and / or a light chain amino acid sequence of SEQ ID NO: 433, (vvv) a heavy chain amino acid sequence of SEQ ID NO: 379,and / or the light chain amino acid sequence of SEQ ID NO: 434, (www) the heavy chain amino acid sequence of SEQ ID NO: 380, and / or the light chain amino acid sequence of SEQ ID NO: 439, (xxx) the heavy chain amino acid sequence of SEQ ID NO: 380, and / or the light chain amino acid sequence of SEQ ID NO: 435, (yyy) the heavy chain amino acid sequence of SEQ ID NO: 381, and / or the light chain amino acid sequence of SEQ ID NO: 436, (zzz) the heavy chain amino acid sequence of SEQ ID NO: 381, and / or the light chain amino acid sequence of SEQ ID NO: 437, (aaaa) the heavy chain amino acid sequence of SEQ ID NO: 381 and / or the light chain amino acid sequence of SEQ ID NO: 438, (bbbb) the heavy chain amino acid sequence of SEQ ID NO: 381 and / or the light chain amino acid sequence of SEQ ID NO: 439, (cccc) the heavy chain amino acid sequence of SEQ ID NO: 382 and / or the light chain amino acid sequence of SEQ ID NO: 439, (dddd) the heavy chain amino acid sequence of SEQ ID NO: 383 and / or the light chain amino acid sequence of SEQ ID NO: 439, (eeee) the heavy chain amino acid sequence of SEQ ID NO: 384 and / or the light chain amino acid sequence of SEQ ID NO: 439, (ffff) SEQ ID NO: 3 85, and / or the light chain amino acid sequence of SEQ ID NO: 439, (gggg) the heavy chain amino acid sequence of SEQ ID NO: 380, and / or the light chain amino acid sequence of SEQ ID NO: 440, (hhhh) the heavy chain amino acid sequence of SEQ ID NO: 384, and / or the light chain amino acid sequence of SEQ ID NO: 440, (iiii) the heavy chain amino acid sequence of SEQ ID NO: 383, and / or the light chain amino acid sequence of SEQ ID NO: 436, (jjjj) the heavy chain amino acid sequence of SEQ ID NO: 383, and / or the light chain amino acid sequence of SEQ ID NO: 438, (kkkk) the heavy chain amino acid sequence of SEQ ID NO: 385, and / or the light chain amino acid sequence of SEQ ID NO: 436, (llll) the heavy chain amino acid sequence of SEQ ID NO: 385, and / or the light chain amino acid sequence of SEQ ID NO: 438, (mmmm) the heavy chain amino acid sequence of SEQ ID NO: 380, and / or the light chain amino acid sequence of SEQ ID NO: 441, (nnnn) the heavy chain amino acid sequence of SEQ ID NO: 380, and / or the light chain amino acid sequence of SEQ ID NO: 436, (oooo) the heavy chain amino acid sequence of SEQ ID NO: 380, and / or the light chain amino acid sequence of SEQ ID NO: 437, (pppp) the heavy chain amino acid sequence of SEQ ID NO: 380, and / or SEQ ID NO: 4 38 light chain amino acid sequence, (qqqq) the heavy chain amino acid sequence of SEQ ID NO: 381 and / or the light chain amino acid sequence of SEQ ID NO: 435, (rrrr) the heavy chain amino acid sequence of SEQ ID NO: 381 and / or the light chain amino acid sequence of SEQ ID NO: 440, (ssss) the heavy chain amino acid sequence of SEQ ID NO: 381 and / or the light chain amino acid sequence of SEQ ID NO: 441, (tttt) the heavy chain amino acid sequence of SEQ ID NO: 386 and / or the light chain amino acid sequence of SEQ ID NO: 442, (uuuu) the heavy chain amino acid sequence of SEQ ID NO: 387 and / or the light chain amino acid sequence of SEQ ID NO: 431,(vvvv) the heavy chain amino acid sequence of SEQ ID NO: 388 and / or the light chain amino acid sequence of SEQ ID NO: 447, (wwww) the heavy chain amino acid sequence of SEQ ID NO: 388 and / or the light chain amino acid sequence of SEQ ID NO: 443, (xxxx) the heavy chain amino acid sequence of SEQ ID NO: 389 and / or the light chain amino acid sequence of SEQ ID NO: 444, (yyyy) the heavy chain amino acid sequence of SEQ ID NO: 389 and / or the light chain amino acid sequence of SEQ ID NO: 445, (zzzz) the heavy chain amino acid sequence of SEQ ID NO: 389 and / or the light chain amino acid sequence of SEQ ID NO: 446, (aaaaa) the heavy chain amino acid sequence of SEQ ID NO: 390 and / or the light chain amino acid sequence of SEQ ID NO: 447, (bbbbb) the heavy chain amino acid sequence of SEQ ID NO: 389 and / or the light chain amino acid sequence of SEQ ID NO: 447, (ccccc) the heavy chain amino acid sequence of SEQ ID NO: 391 and / or the light chain amino acid sequence of SEQ ID NO: 447, (ddddd) the heavy chain amino acid sequence of SEQ ID NO: 392 and / or the light chain amino acid sequence of SEQ ID NO: 447, (eeeee) the heavy chain amino acid sequence of SEQ ID NO: 393 and / or the light chain amino acid sequence of SEQ ID NO: 447, (fffff ) the heavy chain amino acid sequence of SEQ ID NO: 394 and / or the light chain amino acid sequence of SEQ ID NO: 447, (ggggg) the heavy chain amino acid sequence of SEQ ID NO: 390 and / or the light chain amino acid sequence of SEQ ID NO: 448, (hhhhh) the heavy chain amino acid sequence of SEQ ID NO: 388 and / or the light chain amino acid sequence of SEQ ID NO: 448, (iiiiii) the heavy chain amino acid sequence of SEQ ID NO: 391 and / or the light chain amino acid sequence of SEQ ID NO: 448, (jjjjj) the heavy chain amino acid sequence of SEQ ID NO: 392 and / or the light chain amino acid sequence of SEQ ID NO: 448, (kkkkk) SEQ ID NO: 3 93 and / or the light chain amino acid sequence of SEQ ID NO: 448, (lllll) the heavy chain amino acid sequence of SEQ ID NO: 394 and / or the light chain amino acid sequence of SEQ ID NO: 448, (mmmmm) the heavy chain amino acid sequence of SEQ ID NO: 392 and / or the light chain amino acid sequence of SEQ ID NO: 444, (nnnnn) the heavy chain amino acid sequence of SEQ ID NO: 392 and / or the light chain amino acid sequence of SEQ ID NO: 446, (ooooo) the heavy chain amino acid sequence of SEQ ID NO: 394 and / or the light chain amino acid sequence of SEQ ID NO: 444, (ppppp) the heavy chain amino acid sequence of SEQ ID NO: 394 amino acid sequence of SEQ ID NO: 395 and / or the light chain amino acid sequence of SEQ ID NO: 448, (rrrrr) the heavy chain amino acid sequence of SEQ ID NO: 396 and / or the light chain amino acid sequence of SEQ ID NO: 448, (sssss) the heavy chain amino acid sequence of SEQ ID NO: 388 and / or the light chain amino acid sequence of SEQ ID NO: 449, (ttttt) the heavy chain amino acid sequence of SEQ ID NO: 394 and / or the light chain amino acid sequence of SEQ ID NO: 450, (uuuuu) the heavy chain amino acid sequence of SEQ ID NO: 394,and / or the light chain amino acid sequence of SEQ ID NO: 451, (vvvvv) the heavy chain amino acid sequence of SEQ ID NO: 395, and / or the light chain amino acid sequence of SEQ ID NO: 450, (ww) the heavy chain amino acid sequence of SEQ ID NO: 396, and / or the light chain amino acid sequence of SEQ ID NO: 451, (xxxxx) the heavy chain amino acid sequence of SEQ ID NO: 395, and / or the light chain amino acid sequence of SEQ ID NO: 451, (yyyyy) the heavy chain amino acid sequence of SEQ ID NO: 396, and / or the light chain amino acid sequence of SEQ ID NO: 450, (zzzzz) the heavy chain amino acid sequence of SEQ ID NO: 397, and / or the light chain amino acid sequence of SEQ ID NO: 450, (aaaaaa) the heavy chain amino acid sequence of SEQ ID NO: 397 and / or the light chain amino acid sequence of SEQ ID NO: 451, (bbbbbb) the heavy chain amino acid sequence of SEQ ID NO: 395 and / or the light chain amino acid sequence of SEQ ID NO: 452, (cccccc) the heavy chain amino acid sequence of SEQ ID NO: 396 and / or the light chain amino acid sequence of SEQ ID NO: 452, (dddddd) the heavy chain amino acid sequence of SEQ ID NO: 388 and / or the light chain amino acid sequence of SEQ ID NO: 444, (eeeeee) the heavy chain amino acid sequence of SEQ ID NO: 397 and / or the light chain amino acid sequence of SEQ ID NO: 452, (ffffff) the heavy chain amino acid sequence of SEQ ID NO: 397 and / or the light chain amino acid sequence of SEQ ID NO: 448, (gggggg) the heavy chain amino acid sequence of SEQ ID NO: 394 and / or the light chain amino acid sequence of SEQ ID NO: 452, (hhhhhh) the heavy chain amino acid sequence of SEQ ID NO: 398 and / or the light chain amino acid sequence of SEQ ID NO: 444, (iiiiii) the heavy chain amino acid sequence of SEQ ID NO: 399 and / or the light chain amino acid sequence of SEQ ID NO: 444, (jjjjjj) the heavy chain amino acid sequence of SEQ ID NO: 400 and / or the light chain amino acid sequence of SEQ ID NO: 444, (kkkkkk) the heavy chain amino acid sequence of SEQ ID NO: 398 and / or the light chain amino acid sequence of SEQ ID NO: 453, (llllll) the heavy chain amino acid sequence of SEQ ID NO: 399 and / or the light chain amino acid sequence of SEQ ID NO: 453, (mmmmmm) the heavy chain amino acid sequence of SEQ ID NO: 400 and / or the light chain amino acid sequence of SEQ ID NO: 453, (nnnnnn) the heavy chain amino acid sequence of SEQ ID NO: 398 and / or the light chain amino acid sequence of SEQ ID NO: 454, (oooooo) the heavy chain amino acid sequence of SEQ ID NO: 388 and / or the light chain amino acid sequence of SEQ ID NO: 445, (pppppp) the heavy chain amino acid sequence of SEQ ID NO: 399 and / or the light chain amino acid sequence of SEQ ID NO: 454, (qqqqqq) the heavy chain amino acid sequence of SEQ ID NO: 400 and / or the light chain amino acid sequence of SEQ ID NO: 454, (rrrrrr) the heavy chain amino acid sequence of SEQ ID NO: 398 and / or the light chain amino acid sequence of SEQ ID NO: 455, (ssssss) the heavy chain amino acid sequence of SEQ ID NO: 399 and / or the light chain amino acid sequence of SEQ ID NO: 455, (tttttt) the heavy chain amino acid sequence of SEQ ID NO: 400 and / or the light chain amino acid sequence of SEQ ID NO: 455,(uuuuuu) the heavy chain amino acid sequence of SEQ ID NO: 401 and / or the light chain amino acid sequence of SEQ ID NO: 448, (vvvvvv) the heavy chain amino acid sequence of SEQ ID NO: 402 and / or the light chain amino acid sequence of SEQ ID NO: 448, (wwwwww) the heavy chain amino acid sequence of SEQ ID NO: 403 and / or the light chain amino acid sequence of SEQ ID NO: 448, (xxxxxx) the heavy chain amino acid sequence of SEQ ID NO: 401 and / or the light chain amino acid sequence of SEQ ID NO: 451, (yyyyyy) the heavy chain amino acid sequence of SEQ ID NO: 402 and / or the light chain amino acid sequence of SEQ ID NO: 451, (zzzzzz) the heavy chain amino acid sequence of SEQ ID NO: 388 and / or the light chain amino acid sequence of SEQ ID NO: 446, (aaaaaaa) the heavy chain amino acid sequence of SEQ ID NO: 403 and / or the light chain amino acid sequence of SEQ ID NO: 451, (bbbbbbb) the heavy chain amino acid sequence of SEQ ID NO: 401 and / or the light chain amino acid sequence of SEQ ID NO: 450, (ccccccc) the heavy chain amino acid sequence of SEQ ID NO: 402 and / or the light chain amino acid sequence of SEQ ID NO: 450, (ddddddd) the heavy chain amino acid sequence of SEQ ID NO: 403 and / or the light chain amino acid sequence of SEQ ID NO: 450, (eeeeeee) the heavy chain amino acid sequence of SEQ ID NO: 401, and / or the light chain amino acid sequence of SEQ ID NO: 452, (fffffff) the heavy chain amino acid sequence of SEQ ID NO: 402, and / or the light chain amino acid sequence of SEQ ID NO: 452, (ggggggg) the heavy chain amino acid sequence of SEQ ID NO: 403, and / or the light chain amino acid sequence of SEQ ID NO: 452, (hhhhhhh) the heavy chain amino acid sequence of SEQ ID NO: 389, and / or the light chain amino acid sequence of SEQ ID NO: 453, (iiiiiii) the heavy chain amino acid sequence of SEQ ID NO: 389, and / or the light chain amino acid sequence of SEQ ID NO: 454, (j jjjjjj) the heavy chain amino acid sequence of SEQ ID NO: 389 and / or the light chain amino acid sequence of SEQ ID NO: 455, (kkkkkkk) the heavy chain amino acid sequence of SEQ ID NO: 389 and / or the light chain amino acid sequence of SEQ ID NO: 443, (lllllll) the heavy chain amino acid sequence of SEQ ID NO: 391 and / or the light chain amino acid sequence of SEQ ID NO: 451, (mmmmmmm) the heavy chain amino acid sequence of SEQ ID NO: 391 and / or the light chain amino acid sequence of SEQ ID NO: 450, (nnnnnnn) the heavy chain amino acid sequence of SEQ ID NO: 391, and and / or the light chain amino acid sequence of SEQ ID NO: 452, (ooooooo) the heavy chain amino acid sequence of SEQ ID NO: 389 and / or the light chain amino acid sequence of SEQ ID NO: 448, (ppppppp) the heavy chain amino acid sequence of SEQ ID NO: 389 and / or the light chain amino acid sequence of SEQ ID NO: 449, (qqqqqqq) the heavy chain amino acid sequence of SEQ ID NO: 463 and / or the light chain amino acid sequence of SEQ ID NO: 464, or (rrrrrrr) the heavy chain amino acid sequence of SEQ ID NO: 460 and / or the light chain amino acid sequence of SEQ ID NO: 404.

[0013] In one embodiment, the antigen-binding fragment is a Fab, scFab, Fab', F(ab')2, Fv, scFv, diabody, or triabody.

[0014] In another aspect, the present disclosure provides a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof disclosed herein and a pharmaceutically acceptable carrier.

[0015] The present disclosure is also directed to a method of treating congenital adrenal hyperplasia (CAH) in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to CRH. In one embodiment, the present disclosure is directed to a method of treating CAH in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding fragment thereof disclosed herein.

[0016] The present disclosure is also directed to a method of treating CAH in a subject in need thereof, comprising administering to the subject (i) a mineralocorticoid and / or a glucocorticoid, and (ii) an antibody or antigen-binding fragment thereof that specifically binds to CRH. The present disclosure is also directed to a method of treating CAH in a subject in need thereof, comprising administering to the subject (i) a mineralocorticoid and / or a glucocorticoid, and (ii) an antibody or antigen-binding fragment thereof disclosed herein.

[0017] The present disclosure is also directed to a method of reducing androgen levels in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding fragment thereof disclosed herein.

[0018] The present disclosure is also directed to a method of reducing androgen levels in a subject with CAH, comprising administering to the subject (i) a mineralocorticoid and / or a glucocorticoid, and (ii) an antibody or antigen-binding fragment thereof that specifically binds to CRH. The present disclosure is also directed to a method of reducing androgen levels in a subject with CAH, comprising administering to the subject an antibody or antigen-binding fragment thereof disclosed herein.

[0019] The present disclosure is also directed to a method of reducing androgen levels in a subject with CAH, comprising administering to the subject (i) a mineralocorticoid and / or a glucocorticoid, and (ii) an antibody or antigen-binding fragment thereof disclosed herein.

[0020] In one embodiment, the androgen is testosterone, free testosterone, androstenedione, an 11-oxygenated androgen, or a combination thereof. In another embodiment, the 11-oxygenated androgen is 11-hydroxyandrostenedione (11OHA4), 11-hydroxytestosterone (11OHT), 11-ketoandrostenedione (11KA4), 11-ketotestosterone (11KT), or a combination thereof.

[0021] The present disclosure is also directed to a method of reducing the severity of one or more symptoms or signs in a subject having CAH, comprising administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to CRH. The present disclosure is also directed to a method of reducing the severity of one or more symptoms or signs in a subject having CAH, comprising administering to the subject an antibody or antigen-binding fragment thereof disclosed herein.

[0022] The present disclosure is also directed to a method of reducing the severity of one or more symptoms or signs in a subject having CAH, comprising administering to the subject (i) a mineralocorticoid and / or a glucocorticoid, and (ii) an antibody or antigen-binding fragment thereof that specifically binds to CRH. The present disclosure is also directed to a method of reducing the severity of one or more symptoms or signs in a subject having CAH, comprising administering to the subject (i) a mineralocorticoid and / or a glucocorticoid, and (ii) an antibody or antigen-binding fragment thereof disclosed herein. In one embodiment, the one or more symptoms or signs are virilization, acne, oily skin or hair, advanced bone age, precocious puberty, short stature, hirsutism, male pattern baldness, enlarged thyroid cartilage, fused labia minora, fused labia majora, hyperpigmentation of the labia majora or minora, rugation of the labia majora, clitoral megaly, testicular adrenal rest tumor, fertility problems, or irregular or absent menstrual periods.

[0023] The present disclosure is also directed to a method of treating CAH in a subject in need thereof, comprising administering to the subject (i) a glucocorticoid, and (ii) an antibody or antigen-binding fragment thereof that specifically binds to CRH, wherein the dose of the administered glucocorticoid is reduced compared to glucocorticoid monotherapy.The present disclosure is also directed to a method of treating CAH in a subject in need thereof, comprising administering to the subject (i) a glucocorticoid, and (ii) an antibody or antigen-binding fragment thereof disclosed herein, wherein the dose of the administered glucocorticoid is reduced compared to glucocorticoid monotherapy.

[0024] The present disclosure is also directed to a method of treating CAH in a subject in need thereof, comprising administering to the subject (i) a glucocorticoid and (ii) an antibody or antigen-binding fragment thereof that specifically binds to CRH, wherein the dose of the administered glucocorticoid is reduced compared to a combination therapy comprising a glucocorticoid and a small molecule inhibitor of CRH. The present disclosure is also directed to a method of treating CAH in a subject in need thereof, comprising administering to the subject (i) a glucocorticoid and (ii) an antibody or antigen-binding fragment thereof disclosed herein, wherein the dose of the administered glucocorticoid is reduced compared to a combination therapy comprising a glucocorticoid and a small molecule inhibitor of CRH. In one embodiment, the severity of one or more undesirable side effects of the glucocorticoid is reduced compared to glucocorticoid monotherapy. In another embodiment, the severity of one or more undesirable side effects of the glucocorticoid is reduced compared to a combination therapy comprising a glucocorticoid and a small molecule inhibitor of CRH. In another embodiment, the one or more undesirable side effects are osteoporosis, hyperglycemia, diabetes, dyslipidemia, increased appetite, weight gain, Cushing's syndrome, Cushingoid features, growth suppression, adrenal suppression, gastritis, peptic ulcer, gastrointestinal bleeding, hypertension, mood swings, irritability, anxiety, or infection.

[0025] The present disclosure is also directed to a method of reducing the level of one or more biomarkers of CAH in a subject having CAH, the method comprising administering to the subject an antibody or antigen-binding fragment thereof that specifically binds to CRH.The present disclosure is also directed to a method of reducing the level of one or more biomarkers of CAH in a subject having CAH, the method comprising administering to the subject an antibody or antigen-binding fragment thereof disclosed herein.

[0026] The present disclosure is also directed to a method for reducing the level of one or more biomarkers of CAH in a subject having CAH, the method comprising administering to the subject (i) a mineralocorticoid and / or a glucocorticoid, and (ii) an antibody or antigen-binding fragment thereof that specifically binds to CRH. The present disclosure is also directed to a method for reducing the level of one or more biomarkers of CAH in a subject having CAH, the method comprising administering to the subject (i) a mineralocorticoid and / or a glucocorticoid, and (ii) an antibody or antigen-binding fragment thereof disclosed herein. In one embodiment, the one or more biomarkers of CAH are 17-hydroxyprogesterone (17-OHP), adrenocorticotropic hormone (ACTH), or androstenedione.

[0027] The present disclosure also relates to a method for inhibiting the hypothalamic-pituitary-adrenal (HPA) axis in a subject in need thereof, comprising administering to the subject an antibody or antigen-binding fragment thereof disclosed herein. In one embodiment, the mineralocorticoid is fludrocortisone. In another embodiment, the glucocorticoid is beclomethasone, betamethasone, budesonide, cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisone, or triamcinolone.

[0028] In one embodiment of the present disclosure, the CAH is classic CAH. In another embodiment, the CAH is non-classic CAH.

[0029] In one embodiment of the present disclosure, a method of treatment comprises administering to a subject (i) a glucocorticoid, (ii) an antibody or antigen-binding fragment thereof that specifically binds to CRH, and (iii) a CRHR1 antagonist. In one embodiment, the method comprises administering to a subject (i) a glucocorticoid, (ii) an antibody or antigen-binding fragment thereof disclosed herein, and (iii) a CRHR1 antagonist. In one embodiment, the method comprises administering to a subject a mineralocorticoid. In one embodiment, the method comprises administering to a subject a glucocorticoid. In one embodiment, the method comprises administering to a subject a mineralocorticoid and a glucocorticoid.

[0030] In one embodiment, CAH is associated with a mutation or deletion in the 21-hydroxylase gene (CYP21A2). In another embodiment, CAH is associated with a mutation or deletion in the 11-beta-hydroxylase gene (CYP11B1). In another embodiment, CAH is associated with a mutation or deletion in the 3-beta-hydroxysteroid dehydrogenase gene (HSD3B2).

[0031] In one embodiment, the subject is a human, hi another embodiment, the subject is male or female.

[0032] Some aspects of the present invention are herein described, by way of example only, with reference to the accompanying drawings, in which: and in which particular reference is now made in detail to the drawings, it is emphasized that the items shown are by way of example and for the purpose of illustrative discussion of aspects of the invention. [Brief explanation of the drawings]

[0033] [Figure 1] FIG. 1 shows the coding sequence open reading frame of mouse CRH (GenBank NM_205769) along with the CRISPR targeting gRNA and PCR primers used in Example 1. [Figure 2]Figures 2A-2B show PCR confirmation of CRH knockout founder mice. The expected PCR DNA fragment sizes for wild-type mice are 146 bp and 103 bp. PCR fragments smaller than these expected sizes indicated deletions in the founder mice. PCR primers CRHbp70F and CRHbp215R, which flank gRNA1, were used to amplify the expected size of 146 bp from wild-type H2L2 Harbour Mice® (Figure 2A, left panel). PCR primers CRHbp208F and CRHbp310R were used to amplify the expected size of 103 bp from wild-type H2L2 Harbour Mice® (Figure 2A, center panel). PCR primers CRHbp70F and CRHbp310R resulted in a PCR fragment size of 241 bp (Figure 2A, right panel). PCR products smaller than the expected size indicated mutations. Founder mice 1, 2, 4, and 5 were confirmed to carry the mutation. Founder mouse 5 from litter 12198 in Figure 2B carried an 11 bp deletion (5'-CAGCCGGTTCT-3') (SEQ ID NO: 465) and was used for subsequent breeding. [Figure 3] Figure 3 shows the location of an 11-bp deletion (5'-CAGCCGGTTCT-3') (SEQ ID NO: 465) in the CRH gene coding region 157-167, which leads to a downstream reading frameshift and a premature stop codon. This defective CRH open reading frame is unable to synthesize the CRH peptide. Founders with this deletion were selected for further breeding to homozygosity, rescue, and immunization. [Figure 4] Figures 4A-4N show the CRH ELISA binding activity of the indicated antibodies in terms of PR numbers of anti-CRH mAbs (Figures 4A, 4B, 4D, 4E, 4G, 4H, 4J, 4K, 4L, and 4M), as well as the luciferase reporter blocking activity of human CRHR1 (hCRHR1) (Figures 4C, 4F, 4I, and 4N). An irrelevant human IgG1 was used as a negative control. [Figure 5]Figures 5A-5L show the CRH ELISA binding activity of series 1 mAb PR004290 and its ΔPTM variants (Figures 5A-5D), as well as the luciferase reporter blocking activity of human CRHR1 (hCRHR1), human CRHR2 (hCRHR2), mouse CRHR1 (mCRHR1), and mouse CRHR2 (mCRHR2) (Figures 5E-5L). [Figure 6] 6A-6D show the luciferase reporter blocking activity of series 2 mAb PR006669 and its ΔPTM mutants for human CRHR1, human CRHR2, mouse CRHR1, and mouse CRHR2. [Figure 7] Figures 7A-7L show the CRH ELISA binding activity of series 5 mAb PR301777 and its humanized variants (Figures 7A-7D), and the luciferase reporter blocking activity of human CRHR1, human CRHR2, mouse CRHR1, and mouse CRHR2 (Figures 7E-7L). [Figure 8] 8A-8D show the human CRHR1 luciferase reporter blocking activity of series 7 mAb PR302309 and its humanized variants. [Figure 9] 9A-9F show the luciferase reporter blocking activity of series 9 mAb PR302334 and its humanized variants for human CRHR1, human CRHR2, mouse CRHR1, and mouse CRHR2. [Figure 10] 10A-10G show the luciferase reporter blocking activity of series 10 mAb PR302341 and its humanized variants for human CRHR1, human CRHR2, mouse CRHR1, and mouse CRHR2. [Figure 11] 11A-11L show the luciferase reporter blocking activity of series 10 humanized mAbs PR302341-10, PR302341-20, and PR302341-23, and their ΔPTM mutants, for human CRHR1, human CRHR2, mouse CRHR1, and mouse CRHR2. [Figure 12]12A-12B show by ELISA that several anti-CRH mAbs bind to human UCN1, but not to human UCN2 or human UCN3. [Figure 13] Figures 13A-13D show ELISA binding of selected mAbs to CRH and human UCN1. [Figure 14] 14A-14H show human UCN1-mediated inhibition of human CRHR1 and CRHR2 reporter activity. [Figure 15] 15A-15D show that anti-CRH mAb binds to the N-terminal region of the CRH peptide. [Figure 16] FIG. 16 shows a diagram of an epitope binding experiment using the Octet platform. [Figure 17] FIG. 17 shows that anti-CRH mAb PR302050 reduces restraint stress-induced ACTH and corticosterone levels in wild-type mice. [Figure 18] FIG. 18 shows that anti-CRH mAb PR302050 reduces constitutively high ACTH in Mrap1 knockout mice. [Figure 19] 19A-19C show that anti-CRH mAb reduces constitutively high ACTH in Mrap1 knockout mice. [Figure 20] 20A-20B show additional anti-CRH mAbs that reduce constitutively high ACTH in Mrap1 knockout mice. [Figure 21] Figures 21A-21B show that anti-CRH mAbs PR302038, PR005660, and PR302050 dose-dependently reduce plasma ACTH concentrations in Mrap1KO mice at antibody doses of 20 mg / kg and 5 mg / kg. Figure 21A shows results from day 2 after mAb administration. Figure 21B shows results from day 16 after mAb administration. [Figure 22]Figures 22A-22B show that the anti-CRH mAbs PR302334-24 and PR005660 significantly reduced plasma ACTH concentrations in Mrap1KO mice at 20 mg / kg, whereas the anti-CRH mAb PR301429, which has comparable binding affinity but is a nonblocker in vitro, did not inhibit ACTH production. PR303394, an analog of CTRND05, was used as a comparator. Figure 22A shows the results from day 2 after mAb administration. Figure 22B shows the results from day 16 after mAb administration. [Figure 23] Figures 23A-23B show that the anti-CRH mAb PR3024064 is more effective than SSR125543A at inhibiting restraint-induced ACTH (Figure 23A) and corticosterone (Figure 23B) release. [Figure 24] Figures 24A-24B show that the anti-CRH mAbs PR302064 and PR302334-24 had superior efficacy to SSR125543A in a model of constitutively elevated ACTH in Mrap1KO mice. ACTH levels are shown on days 2 (Figure 24A) and 14 (Figure 24B) after mAb administration. [Figure 25] Figures 25A-25B show that the anti-CRH mAb PR302334-24 had superior efficacy to SSR125543A in the wild-type mouse restraint stress-induced high ACTH and corticosterone model. Induced plasma levels of ACTH (Figure 25A) and corticosterone (Figure 25B) after mAb administration are shown. [Figure 26] Figure 26A is a diagram of the method for testing the pharmacokinetics of anti-CRH mAb. Specifically, anti-CRH mAb in human IgG1WT-Fc present in mouse serum was captured with a goat anti-human Fc polyclonal antibody and then detected with a goat anti-human (H+L) secondary antibody conjugated to HRP. Figures 26B-26D show the mean serum concentration-time profiles of anti-CRH mAb administered at a single intravenous dose of 5 mg / kg to female C57BL / 6 wild-type mice. DETAILED DESCRIPTION OF THE INVENTION

[0034] Various terms relating to aspects of the present disclosure are used throughout the specification and claims. Such terms are to be given their ordinary meaning in the art unless otherwise indicated. Other specifically defined terms are to be interpreted consistently with the definitions provided herein.

[0035] 1.Definition The term "antibody" (Ab) includes, but is not limited to, glycoprotein immunoglobulins that specifically bind to an antigen and contain at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each H chain contains a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region contains three constant domains, CH1, CH2, and CH3. Each light chain contains a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region contains one constant domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability called complementarity-determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each VH and VL contains three CDRs and four FRs, arranged from amino terminus to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant regions of antibodies can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. The heavy chain may or may not have a C-terminal lysine. Unless otherwise specified herein, amino acids in the variable regions are numbered using the Kabat numbering system, and amino acids in the constant regions are numbered using the EU system.

[0036] The term "monoclonal antibody," as used herein, refers to an antibody produced by a single clone of B cells and binding to the same epitope. In contrast, the term "polyclonal antibody" refers to a population of antibodies produced by different B cells and binding to different epitopes of the same antigen. The term "antibody" includes, by way of example, monoclonal and polyclonal antibodies, chimeric and humanized antibodies, human or non-human antibodies, fully synthetic antibodies, and single-chain antibodies. Non-human antibodies can be humanized by recombinant methods to reduce immunogenicity in humans.

[0037] An antibody may be one that has been modified (e.g., by mutation, deletion, substitution, conjugation to a non-antibody moiety). For example, an antibody may contain one or more variant amino acids that alter the properties (e.g., functional properties) of the antibody (compared to the naturally occurring antibody). For example, many such modifications are known in the art and affect, for example, the half-life, effector functions, and / or immune response of the antibody in a patient. The term antibody also includes artificial polypeptide constructs that contain at least one antibody-derived antigen-binding site.

[0038] An "antigen-binding fragment" of an antibody refers to one or more fragments or portions of an antibody that retain the ability to specifically bind to the antigen bound by the full-length antibody. It has been shown that the antigen-binding function of an antibody can be performed by fragments or portions of a full-length antibody. Examples of binding fragments encompassed within the term "antigen-binding portion" or "antigen-binding fragment" of an antibody described herein include the following: The term "antibody fragment" refers to a portion of an intact antibody. An "antigen-binding fragment," "antigen-binding domain," or "antigen-binding region" refers to a portion of an intact antibody that binds to an antigen. An antigen-binding fragment can contain an antigen-determining region of an intact antibody (e.g., a complementarity-determining region (CDR)). Examples of antigen-binding fragments of antibodies include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments, linear antibodies, and single-chain antibodies. Antigen-binding fragments of antibodies can be derived from any animal species, such as rodents (e.g., mice, rats, or hamsters) and humans, or can be artificially produced.

[0039] Furthermore, the two domains of an Fv fragment, VL and VH, are encoded by separate genes but can be linked using recombinant methods by a synthetic linker that allows the pair of VL and VH regions to be produced as a single protein chain forming a monovalent molecule (known as single-chain Fvs (scFVs); see, e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single-chain antibodies are also intended to be encompassed within the term "antigen-binding portion" or "antigen-binding fragment" of an antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for utility in the same manner as intact antibodies. Antigen-binding portions can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact immunoglobulins. In some embodiments, an antibody is an antigen-binding fragment.

[0040] As used herein, the terms "variable region" and "variable domain" are interchangeable and are common in the art. A variable region typically refers to a portion of an antibody, generally a portion of either the light or heavy chain, typically about the amino-terminal 110-120 or 110-125 amino acids of the mature heavy chain and about 90-115 amino acids of the mature light chain, which vary extensively in sequence among antibodies and are used to determine the binding and specificity of a particular antibody to a particular antigen. Sequence variability is concentrated in regions called complementarity-determining regions (CDRs), while more highly conserved regions in variable domains are called framework regions (FRs). While not wishing to be bound by any particular mechanism or theory, it is believed that the CDRs of the light and heavy chains are primarily responsible for the interaction and specificity of an antibody with an antigen. In some embodiments, the variable region is a mammalian variable region, e.g., a human or rabbit variable region. In some embodiments, the variable region comprises rodent or murine CDRs and human framework regions (FRs). In some embodiments, the variable region is a primate (e.g., non-human primate) variable region. In some embodiments, the variable region comprises rodent or murine CDRs and primate (e.g., non-human primate) framework regions (FRs).

[0041] The term "complementarity determining region" or "CDR" as used herein refers to each region of an antibody variable domain that is hypervariable in sequence and / or forms structurally defined loops (hypervariable loops) and / or contains antigen-contacting residues. An antibody can contain six CDRs, e.g., three in the VH and three in the VL.

[0042] The terms "VL" and "VL domain" are used interchangeably to refer to the light chain variable region of an antibody.

[0043] The terms "VH" and "VH domain" are used interchangeably to refer to the heavy chain variable region of an antibody.

[0044] Terms such as "Kabat numbering" are art-recognized and refer to a system for numbering amino acid residues in the heavy and light chain variable regions of an antibody or antigen-binding fragment thereof. In some embodiments, CDRs can be determined according to the Kabat numbering system (see, e.g., Kabat EA & Wu TT (1971) Ann NY Acad Sci 190: 382-391 and Kabat EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242). Using the Kabat numbering system, the CDRs within an antibody heavy chain molecule are typically located at amino acid positions 31-35 (which may optionally include one or two additional amino acids after 35, designated 35A and 35B according to the Kabat numbering scheme) (CDR1), amino acid positions 50-65 (CDR2), and amino acid positions 95-102 (CDR3). Using the Kabat numbering system, the CDRs within an antibody light chain molecule are typically located at amino acid positions 24-34 (CDR1), amino acid positions 50-56 (CDR2), and amino acid positions 89-97 (CDR3).

[0045] Chothia, instead, refers to the location of the structural loops (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). When numbered using the Kabat numbering convention, the end of the Chothia CDR-H1 loop varies between H32 and H34 depending on the length of the loop (this is because the Kabat numbering scheme places insertions at H35A and H35B, so that if neither 35A nor 35B are present, the loop ends at 32; if only 35A is present, the loop ends at 33; and if both 35A and 35B are present, the loop ends at 34). The AbM hypervariable regions represent a compromise between the Kabat CDRs and Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software.

[0046] As used herein, the terms "constant region" or "constant domain" are used interchangeably and have the meaning commonly used in the art. The constant region is a portion of an antibody, e.g., the carboxyl-terminal portion of the light and / or heavy chain, that is not directly involved in binding the antibody to an antigen but can exhibit various effector functions, such as interacting with Fc receptors. The constant region of an immunoglobulin molecule generally has a more conserved amino acid sequence than the immunoglobulin variable domain. In some embodiments, the antigen or antigen-binding fragment comprises a constant region or a portion thereof that is sufficient for antibody-dependent cell-mediated cytotoxicity (ADCC).

[0047] As used herein, the term "heavy chain," when used in reference to an antibody, can refer to any unique type based on the amino acid sequence of the constant domain, e.g., alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), which give rise to antibodies of the IgA, IgD, IgE, IgG, and IgM classes, respectively, including subclasses of IgG, e.g., IgG1, IgG2, IgG3, and IgG4. Heavy chain amino acid sequences are well known in the art. In some embodiments, the heavy chain is a human heavy chain.

[0048] As used herein, the term "light chain," when used in reference to an antibody, can refer to any specific type, e.g., kappa (κ) or lambda (λ), based on the amino acid sequence of the constant domain. Light chain amino acid sequences are well known in the art. In some embodiments, the light chain is a human light chain.

[0049] The term "chimeric" antibody or antigen-binding fragment thereof refers to an antibody or antigen-binding fragment thereof whose amino acid sequence is derived from two or more species. Typically, the variable regions of both the light and heavy chains correspond to the variable regions of an antibody or antigen-binding fragment thereof derived from one species of mammal (e.g., mouse, rat, rabbit, etc.) having the desired specificity, affinity, and capacity, while the constant regions are homologous to the sequences of an antibody or antigen-binding fragment thereof derived from another species (usually human) to avoid eliciting an immune response in that species.

[0050] The term "humanized" antibody or antigen-binding fragment thereof refers to a form of a non-human (e.g., murine) antibody or antigen-binding fragment thereof that is a specific immunoglobulin chain, chimeric immunoglobulin, or fragment thereof that contains minimal non-human (e.g., murine) sequence. Typically, humanized antibodies or antigen-binding fragments thereof are human immunoglobulins in which residues of the complementarity-determining regions (CDRs) have been replaced with residues from CDRs of a non-human species (e.g., mouse, rat, rabbit, hamster) having the desired specificity, affinity, and capacity ("CDR-grafted") (Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-327 (1988); Verhoeyen et al., Science 239:1534-1536 (1988)). In some cases, Fv framework region (FR) residues of a human immunoglobulin are replaced with corresponding residues in an antibody or fragment of a non-human species having the desired specificity, affinity, and capacity. Humanized antibodies or antigen-binding fragments thereof can be further modified by substitution of additional residues either within the Fv framework regions and / or the replaced non-human residues to refine the specificity, affinity, and / or capacity of the antibody or antigen-binding fragment. Generally, a humanized antibody or antigen-binding fragment thereof will comprise substantially all of at least one, typically two or three, variable domains containing all or substantially all of the CDR regions corresponding to a non-human immunoglobulin, while all or substantially all of the FR regions are of a human immunoglobulin consensus sequence. A humanized antibody or antigen-binding fragment thereof can also comprise at least a portion of an immunoglobulin constant region or domain (Fc), typically of a human immunoglobulin.Examples of methods used to generate humanized antibodies are described in U.S. Patent No. 5,225,539, Roguska et al., Proc. Natl. Acad. Sci., USA, 91(3):969-973 (1994), and Roguska et al., Protein Eng. 9(10):895-904 (1996). In some embodiments, a "humanized antibody" is a resurfaced antibody.

[0051] The term "human" antibody or antigen-binding fragment thereof means an antibody or antigen-binding fragment thereof having an amino acid sequence derived from the human immunoglobulin locus, and such an antibody or antigen-binding fragment thereof is made using any technique known in the art. This definition of human antibody or antigen-binding fragment thereof includes intact or full-length antibodies and fragments thereof.

[0052] "Binding affinity" generally refers to the strength of the total non-covalent interactions between a single binding site of a molecule (e.g., an antibody or antigen-binding fragment thereof) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody or antigen-binding fragment thereof and an antigen). The affinity of a molecule X to its partner Y can generally be expressed by a dissociation constant (KD). Affinity can be measured and / or expressed by a number of methods known in the art, including, but not limited to, the equilibrium dissociation constant (KD) and the equilibrium association constant (KA). KD is determined by the k off / k on KA is calculated by the quotient of k on / k off It is calculated as the quotient of k on refers to, for example, the binding rate constant of an antibody or antigen-binding fragment thereof to an antigen, and k off k refers, for example, to the dissociation of an antibody or antigen-binding fragment thereof from an antigen. on and koff can be determined by techniques known to those skilled in the art, such as Octet BLI, BIAcore® or KinExA.

[0053] As used herein, "epitope" is a term of the art and refers to a localized region of an antigen to which an antibody or antigen-binding fragment thereof can specifically bind. An epitope can be, for example, consecutive amino acids of a polypeptide (a linear or continuous epitope), or an epitope can be, for example, two or more non-contiguous regions of a polypeptide taken together (a conformational, non-linear, discontinuous or discontinuous epitope). In some embodiments, the epitope to which an antibody or antigen-binding fragment thereof binds can be determined by, for example, NMR spectroscopy, X-ray diffraction crystallography studies, ELISA assays, hydrogen / deuterium exchange coupled with mass spectrometry (e.g., liquid chromatography electrospray mass spectrometry), array-based oligopeptide scanning assays, and / or mutagenesis mapping (e.g., site-directed mutagenesis mapping). In X-ray crystallography, crystallization can be achieved using any method known in the art (e.g., Giege R et al., (1994) Acta Crystallogr D Biol Crystallogr 50(Pt 4): 339-350; McPherson A (1990) Eur J Biochem 189: 1-23; Chayen NE (1997) Structure 5: 1269-1274; McPherson A (1976) J Biol Chem 251: 6300-6303).Antibody / antigen-binding fragment thereof:antigen crystals can be studied using well-known X-ray diffraction techniques and refined using computer software such as X-PLOR (Yale University, 1992, distributed by Molecular Simulations, Inc., see e.g., Meth Enzymol (1985) volumes 114 & 115, eds. Wyckoff HW et al.; US 2004 / 0014194) and BUSTER (Bricogne G (1993) Acta Crystallogr D Biol Crystallogr 49(Pt 1): 37-60; Bricogne G (1997) Meth Enzymol 276A: 361-423, ed. Carter CW; Roversi P et al., (2000) Acta Crystallogr D Biol Crystallogr 56(Pt 10): 1316-1323). Mutagenesis mapping studies can be accomplished using any method known to those skilled in the art. For example, for a description of mutagenesis techniques, including alanine scanning mutagenesis techniques, see Champe M et al., (1995) J Biol Chem 270: 1388-1394, and Cunningham BC & Wells JA (1989) Science 244: 1081-1085.

[0054] An "isolated" polypeptide, antibody, polynucleotide, vector, cell, or composition is a polypeptide, antibody, polynucleotide, vector, cell, or composition in a form not found in nature. Isolated polypeptides, antibodies, polynucleotides, vectors, cells, or compositions include those that have been purified to the extent that they are not in a form found in nature. In some embodiments, an isolated antibody, polynucleotide, vector, cell, or composition is substantially pure. As used herein, "substantially pure" refers to a material that is at least 50% pure (i.e., free from contaminants), at least 90% pure, at least 95% pure, at least 98% pure, or at least 99% pure.

[0055] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymers of amino acids of any length. The polymers may be linear or branched, may comprise modified amino acids, and may be interrupted by non-amino acids. The term also encompasses amino acid polymers that are modified naturally or by intervention, for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, such as conjugation with a labeling component. Also included within the definition are polypeptides containing, for example, one or more analogs of an amino acid (including, for example, unnatural amino acids), as well as other modifications known in the art. Because the polypeptides of the present invention are based on antibodies, it is understood that in some embodiments the polypeptides can occur as single chains or associated chains.

[0056] As used herein, the term "host cell" can be any type of cell, for example, a primary cell, a cell in culture, or a cell line. In some embodiments, the term "host cell" refers to a cell transfected with a nucleic acid molecule and the progeny or potential progeny of such a cell. The progeny of such a cell may not be identical to the parent cell transfected with the nucleic acid molecule due, for example, to mutations or environmental influences that may occur in subsequent generations or that may occur upon integration of the nucleic acid molecule into the host cell genome.

[0057] The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes any solvents, cosolvents, complexing agents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, which are not biologically or otherwise undesirable. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in therapeutic formulations is contemplated. Supplementary active ingredients can also be incorporated into the formulation. In addition, various excipients, such as those commonly used in the art, can be included. These and other such compounds are described in the literature, for example, in The Merck Index, Merck & Company, Rahway, NJ. Considerations for the inclusion of various ingredients in pharmaceutical compositions are described, for example, in Gilman et al. (Eds.) (2010); Goodman and Gilman's: The Pharmacological Basis of Therapeutics, 12th Ed., The McGraw-Hill Companies.

[0058] "Subject," as used herein, means a human or non-human mammal, such as a dog, cat, mouse, rat, cow, sheep, pig, goat, non-human primate, or bird, such as a chicken, as well as any other vertebrate or invertebrate. In some embodiments, the subject is a human.

[0059] In some embodiments, the subject is experiencing and / or exhibiting at least one symptom of the disease or disorder to be treated and / or prevented. In some embodiments, the subject has been confirmed or diagnosed with congenital adrenal hyperplasia (CAH). In some embodiments, the subject is suspected of having CAH. In some embodiments, the subject has clinical records indicating that the subject has CAH (the clinical records may indicate that the subject should be treated with the antibody or antigen-binding fragment thereof provided herein).

[0060] As used herein, the term "treat" or "treatment" refers to curative or palliative measures. Beneficial or desirable clinical results include, but are not limited to, the total or partial alleviation of symptoms associated with a disease or disorder or condition, whether detectable or undetectable, a decrease in the extent of the disease, a stable (i.e., not worsening) state of the disease, a delay or slowing of disease progression, an improvement or alleviation of the disease state (e.g., one or more symptoms of the disease), and remission (whether partial or total). "Treatment" can also mean prolonging survival compared to expected survival if not receiving treatment.

[0061] The term "preventing" as used herein means preventing the onset, recurrence or spread, in whole or in part, of a disease or condition described herein, or a symptom thereof.

[0062] As used herein, a "therapeutically effective amount" is an amount of an antibody or antigen-binding fragment thereof disclosed herein that is sufficient to achieve a desired effect, and may vary depending on the nature and severity of the disease state and the potency of the antibody / fragment. The therapeutic effect is some alleviation of one or more symptoms of the disease, and may include curing the disease. "Curing" means that the symptoms of active disease are eliminated. However, certain long-term or permanent effects of the disease may exist even after a cure is achieved (e.g., extensive tissue damage, etc.).

[0063] In some embodiments, the antibodies and antigen-binding fragments described herein include monoclonal antibodies that are antibodies or antigen-binding fragments thereof against corticotropin-releasing hormone (CRH), also known as corticotropin-releasing factor (CRF) or CRH1. CRH is a neuropeptide hormone that activates the synthesis and release of adrenocorticotropic hormone (ACTH) from the pituitary gland. CRH regulates various neuroendocrine, sympathetic, and behavioral functions, including important roles in stress response, anxiety and depression, wakefulness, feeding behavior, energy metabolism, and digestive and cardiovascular function. CRH is a 41-amino acid peptide derived by enzymatic cleavage of a 196-amino acid preprohormone and is secreted from the paraventricular nucleus (PVN) of the hypothalamus. The amino acid sequence of human and mouse CRH (UniProtKB-P06850; Genbank accession number EAW86897.1) is SEEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRKLMEII (SEQ ID NO: 466).

[0064] CRH acts through two unique G protein-coupled receptors, CRHR1 and CRHR2. CRHR1 expression is widespread in brain regions involved in sensory and motor control, such as the cortical pallial, olfactory bulb, hippocampus, amygdala, basal ganglia, medial and lateral hypothalamic nuclei, and cerebellum. In contrast, CRHR2 expression is predominant in subcortical regions, including the lateral septum, bed nucleus of the stria terminalis, ventromedial hypothalamic nucleus, and medial and central nuclei of the amygdala. In the anterior pituitary, CRHR1 mediates ACTH release in response to CRH.

[0065] In response to stress, the hypothalamus releases CRH, which triggers the release of ACTH from the anterior pituitary gland into the circulation. ACTH then binds to receptors in the adrenal cortex, triggering the release of stress hormones such as cortisol. This entire system, known as the hypothalamic-pituitary-adrenal (HPA) axis, plays an important role in regulating the fight-or-flight response to stress.

[0066] The term "small molecule inhibitors of CRH," as used herein, refers to compounds or molecules having a molecular weight of approximately 2000 daltons or less that inhibit corticotropin-releasing hormone (CRH). Small molecule inhibitors of CRH include, but are not limited to, antagonists of CRH receptor 1 (CRHR1), also known as the corticotropin-releasing factor type 1 (CRF1) receptor. More specific examples of inhibitors include, but are not limited to, Crinecerfont (SSR125543A or NBI-74788, e.g., WO 2020 / 115555), Antalarmin (N-butyl-N-ethyl-(2,5,6-trimethyl-7-(2,4,6-trimethylphenyl)-7H-pyrrolo(2,3-d)pyrimidin-4-yl)amine, U.S. Patent Application Publication No. 2017 / 0020877), and Tildacerfont (3-(4-chloro-2-morpholin-4-yl-thiazol-5-yl)-7-(l-ethyl-propyl)-2,5-dimethyl-pyrazolo[1,5-α]pyrimidine, WO 2008 / 036579).

[0067] The use of the alternative (e.g., "or") should be understood to mean either one, both, or any combination of the alternatives. As used herein, the indefinite article "a" or "an" should be understood to refer to "one or more" of any of the listed or arranged components.

[0068] The term "and / or," as used herein, is considered a specific disclosure of each of the two characteristic features or components with or without the other. Thus, when the term "and / or" is used herein in phrases such as "A and / or B," it is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Similarly, when the term "and / or" is used in phrases such as "A, B, and / or C," it is intended to encompass the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone), respectively.

[0069] Where embodiments are described herein with the word "comprising," it is understood that other similar embodiments described with the terms "consisting of" and / or "consisting essentially of" are also provided.

[0070] The term "about" refers to a value or composition that is within an acceptable error range of a particular value or composition as determined by one of ordinary skill in the art, which depends in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, "about" can mean within or more than 1 standard deviation, according to practice in the art. Alternatively, "about" can mean a range of up to 10% or 20% (i.e., ±10% or ±20%). For example, about 3 mg can include any number between 2.7 mg and 3.3 mg (about 10%) or between 2.4 mg and 3.6 mg (about 20%). Furthermore, particularly with respect to biological systems or processes, the term can mean up to an order of magnitude off the value or up to five times the value. When specific values ​​or compositions are provided in this application and claims, unless otherwise stated, the meaning of "about" is presumed to be within an acceptable error range of the particular value or composition.

[0071] As described herein, any concentration range, percentage range, ratio range, or integer range is understood to include any integer value within the recited range, and, where appropriate, fractions thereof (e.g., integer tenths and hundredths), unless otherwise indicated.

[0072] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art and related to this disclosure.For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 5th ed., 2013, Academic Press; and the Oxford Dictionary of Biochemistry and Molecular Biology, 2006, Oxford University Press provide those skilled in the art with a general dictionary for many of the terms used in this disclosure.

[0073] Units, prefixes, and symbols are shown in the form accepted by the Systeme International de Unites (SI). Numerical ranges are inclusive of the numbers defining the range. The headings provided herein are not limitations of the various aspects of this disclosure, which may be by reference to the specification as a whole. Accordingly, the terms defined immediately below are more fully defined by reference to the specification as a whole.

[0074] Various aspects are described in further detail in the following sections.

[0075] 2. Anti-CRH antibody or antigen-binding fragment thereof In some embodiments of the present disclosure, the antibody or antigen-binding fragment thereof described herein specifically binds to human CRH. In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable domain (VH) CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 22-35, a VH CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 53-78, and a VH CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 113-122 or the amino acid sequence of a PDV or GID, and / or a light chain variable domain (VL) CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 152-174, a VL CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 193-198, and a VL CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 223-232.

[0076] In some embodiments, the antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence at least 90%, 95%, or 99% identical to any one of SEQ ID NOs: 240-295, 459, and 461, and / or a VL comprising an amino acid sequence at least 90%, 95%, or 99% identical to any one of SEQ ID NOs: 296-347 and 462. In some embodiments, the antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence of any one of SEQ ID NOs: 240-295, 459, and 461, and / or a VL comprising the amino acid sequence of any one of SEQ ID NOs: 296-347 and 462.

[0077] In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain comprising an amino acid sequence at least 90%, 95%, or 99% identical to any one of SEQ ID NOs: 348-403, 460, and 463, and / or a light chain comprising an amino acid sequence at least 90%, 95%, or 99% identical to any one of SEQ ID NOs: 404-455, and 464. In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain comprising an amino acid sequence at least 90%, 95%, or 99% identical to any one of SEQ ID NOs: 348-403, 460, and 463, and / or a light chain comprising the amino acid sequence of any one of SEQ ID NOs: 404-455, and 464.

[0078] In some embodiments, the antibody or antigen-binding fragment is an antibody or antigen-binding fragment thereof of Table 1.

[0079] [Table 1-1]

[0080] [Table 1-2]

[0081] [Table 1-3]

[0082] [Table 1-4]

[0083] [Table 1-5]

[0084] [Table 1-6]

[0085]

Table 1-7

[0086]

Table 1-8

[0087]

Table 1-9

[0088]

Table 1-10

[0089]

Table 1-11

[0090]

Table 1-12

[0091]

Table 1-13

[0092]

Table 1-14

[0093]

Table 1-15

[0094] In some embodiments, provided herein are polynucleotides comprising nucleotide sequences encoding an antibody or antigen-binding fragment thereof, or a domain thereof (e.g., variable light chain region and / or variable heavy chain region) described herein linked to human CRH and a vector, e.g., a vector comprising the polynucleotide such that it is recombinantly expressed in host cells (e.g., E. coli and mammalian cells).

[0095] In some embodiments, provided herein is a polynucleotide comprising a nucleic acid molecule encoding a VH comprising an amino acid sequence at least 90%, 95%, or 99% identical to any one of SEQ ID NOs: 240-295, and / or a VL comprising an amino acid sequence at least 90%, 95%, or 99% identical to any one of SEQ ID NOs: 296-347. In some embodiments, the polynucleotide encodes an antibody or antigen-binding fragment comprising a VH comprising the amino acid sequence of any one of SEQ ID NOs: 240-295, and / or a VL comprising the amino acid sequence of any one of SEQ ID NOs: 296-347.

[0096] In some embodiments, the polynucleotide encodes an antibody or antigen-binding fragment thereof comprising a heavy chain comprising an amino acid sequence at least 90%, 95%, or 99% identical to any one of SEQ ID NOs: 348-403, and / or a light chain comprising an amino acid sequence at least 90%, 95%, or 99% identical to any one of SEQ ID NOs: 404-455. ...

[0097] Polynucleotides encoding the antibodies or antigen-binding fragments thereof, or domains thereof, described herein can be generated from nucleic acid from a suitable source (e.g., a hybridoma) using methods well known in the art (e.g., PCR and other molecular cloning methods). For example, PCR amplification using synthetic primers hybridizable to the 3' and 5' ends of a known sequence can be performed using genomic DNA obtained from hybridoma cells producing the antibody of interest. Such PCR amplification methods can be used to obtain nucleic acids containing sequences encoding the light and / or heavy chains of the antibodies or antigen-binding fragments thereof described herein. Such PCR amplification methods can be used to obtain nucleic acids containing sequences encoding the variable light chain region and / or variable heavy chain region of the antibodies or antigen-binding fragments thereof. Amplified nucleic acids can be cloned into vectors for expression in host cells and for further cloning, e.g., to generate antibodies or antigen-binding fragments thereof.

[0098] The polynucleotides provided herein may be, for example, in the form of RNA or DNA. DNA includes cDNA, genomic DNA, and synthetic DNA, and the DNA may be double-stranded or single-stranded. If single-stranded, the DNA may be a coding strand or a non-coding (antisense) strand. In some embodiments, the polynucleotide is a cDNA or a DNA lacking one or more endogenous introns. In some embodiments, the polynucleotide is a non-naturally occurring polynucleotide. In some embodiments, the polynucleotide is recombinantly produced. In some embodiments, the polynucleotide is isolated. In some embodiments, the polynucleotide is substantially pure. In some embodiments, the polynucleotide is purified from natural components.

[0099] In some embodiments, provided herein are vectors (e.g., expression vectors) comprising polynucleotides encoding the amino acid sequences disclosed herein. In some embodiments, provided herein are vectors (e.g., expression vectors) comprising polynucleotides encoding the variable heavy chain (VH) complementarity determining regions (CDRs), variable light chain (VL) CDRs, variable heavy chains (VH), variable light chains (VL), heavy chains (HC), and light chains (LC) disclosed herein.

[0100] Also provided herein are cells, eg, host cells, comprising such vectors that recombinantly express an anti-CRH antibody or antigen-binding fragment thereof.

[0101] In some embodiments, provided herein are vectors (eg, expression vectors) comprising a polynucleotide comprising a nucleotide sequence encoding antibodies and antigen-binding fragments thereof that specifically bind to human CRH.

[0102] Methods well known to those skilled in the art can be used to construct expression vectors containing the protein or antibody or antigen-binding fragment thereof or domain (e.g., light or heavy chain) coding sequence and appropriate transcriptional and translational control signals. These methods include, for example, in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. Replicable vectors containing nucleic acid sequences encoding the protein or antibody or antigen-binding fragment thereof, heavy or light chain, heavy or light chain variable domain, or heavy or light chain CDR described herein, operably linked to a promoter, are also provided.

[0103] The expression vector can be transferred into a cell (e.g., a host cell) by conventional techniques, and the resulting cells can then be cultured by conventional techniques to produce a protein described herein, or an antibody or antigen-binding fragment thereof (e.g., an antibody or antigen-binding fragment thereof comprising the six CDRs, VH, VL, VH and VL, heavy chain, light chain, or heavy and light chain of an antibody disclosed herein), or a domain thereof (e.g., the VH, VL, VH and VL, heavy chain, or light chain of an antibody disclosed herein). Thus, provided herein are host cells containing a polynucleotide encoding a protein described herein, or an antibody or antigen-binding fragment thereof (e.g., the six CDRs, VH, VL, VH and VL, heavy chain, light chain, or heavy and light chain of an antibody disclosed herein), or a domain thereof (e.g., the VH, VL, VH and VL, heavy chain, or light chain of an antibody disclosed herein), wherein such sequence is operably linked to a promoter for expression in the host cell. In some embodiments, for expression of a double-chain antibody or antigen-binding fragment thereof, vectors encoding both the heavy and light chains can be individually co-expressed in a host cell to express the entire immunoglobulin. In some embodiments, the host cell contains a vector comprising polynucleotides encoding both the heavy and light chains of an antibody or domain thereof described herein. In some embodiments, the host cell contains two different vectors: a first vector comprising polynucleotides encoding the heavy chain or heavy chain variable region of an antibody or antigen-binding fragment thereof described herein, and a second vector comprising polynucleotides encoding the light chain or light chain variable region of an antibody or antigen-binding fragment thereof described herein. In some embodiments, the first host cell contains a first vector comprising polynucleotides encoding the heavy chain or heavy chain variable region of an antibody or antigen-binding fragment thereof described herein, and the second host cell contains a second vector comprising polynucleotides encoding the light chain or light chain variable region of an antibody or antigen-binding fragment thereof described herein.In some embodiments, the heavy chain / heavy chain variable region expressed by the first cell associates with the light chain / light chain variable region of the second cell to form an antibody or antigen-binding fragment thereof described herein. In some embodiments, provided herein is a population of host cells comprising such a first host cell and such a second host cell.

[0104] In some embodiments, a population of vectors is provided herein, comprising a first vector comprising a polynucleotide encoding the light chain / light chain variable region of an antibody or antigen-binding fragment thereof described herein, and a second vector comprising a polynucleotide encoding the heavy chain / heavy chain variable region of an antibody or antigen-binding fragment thereof described herein. Alternatively, a single vector can be used that encodes and expresses both heavy and light chain polypeptides.

[0105] A variety of host-expression vector systems can be utilized to express the proteins, and antibodies and antigen-binding fragments thereof, described herein. Such host-expression systems represent vehicles by which a coding sequence of interest can be produced and subsequently purified, but also represent cells that, when transformed or transfected with the appropriate nucleotide coding sequence, are capable of expressing the proteins, or antibodies or antigen-binding fragments thereof, described herein in situ. These include microorganisms such as bacteria (e.g., Escherichia coli and B. subtilis) transformed with recombinant bacteriophage DNA, plasmid DNA, or cosmid DNA expression vectors containing antibody coding sequences; yeast (e.g., Saccharomyces pichia) transformed with recombinant yeast expression vectors containing antibody coding sequences; insect cell lines infected with recombinant viral expression vectors (e.g., baculovirus) containing antibody coding sequences; and plant cell lines (e.g., Chlamydomonas reinhardtii) infected with recombinant viral expression vectors (cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing antibody coding sequences. reinhardtii); or mammalian cell lines (e.g., COS (e.g., COS1 or COS), CHO, BHK, MDCK, HEK293, NS0, PER.C6, VERO, CRL7030, HsS78Bst, HeLa, and NIH3T3, HEK-293T, HepG2, SP210, R1.1, BW, LM, BSC1, BSC40, YB / 20, and BMT10 cells) harboring a recombinant expression construct containing a promoter derived from the genome of a mammalian cell (e.g., a metallothionein promoter) or a promoter derived from a mammalian virus (e.g., an adenovirus late promoter, a vaccinia virus 7.5K promoter).In some embodiments, cells expressing the proteins, or antibodies, and antigen-binding fragments thereof, described herein are CHO cells, e.g., CHO cells from the CHO GS System™ (Lonza). In some embodiments, cells expressing the proteins, or antibodies, and antigen-binding fragments thereof, described herein are human cells, e.g., a human cell line. In some embodiments, the mammalian expression vector is pOptiVEC™ or pcDNA3.3. In some embodiments, bacterial cells such as Escherichia coli or eukaryotic cells (e.g., mammalian cells) are used to express recombinant antibody molecules, particularly for the expression of whole recombinant antibody molecules. For example, mammalian cells such as Chinese hamster ovary (CHO) cells in combination with vectors such as the human cytomegalovirus major intermediate-early gene promoter element are effective expression systems for antibodies (Foecking MK & Hofstetter H (1986) Gene 45: 101-105; and Cockett MI et al., (1990) Biotechnology 8: 662-667). In some embodiments, the proteins, or antibodies or antigen-binding fragments thereof described herein are produced by HEK-293T cells.

[0106] In addition, a host cell strain can be chosen that modulates the expression of the inserted sequences, or modifies and processes the gene product in the specific fashion desired. Such modifications (e.g., glycosylation) and processing (e.g., cleavage) of protein products may contribute to the function of the protein. To this end, eukaryotic host cells that possess the cellular machinery for proper processing of the primary transcript, glycosylation, and phosphate of the gene product can be used. Such mammalian cells include, but are not limited to, CHO, VERO, BHK, Hela, MDCK, HEK293, NIH3T3, W138, BT483, Hs578T, HTB2, BT2O and T47D, NS0 (a murine myeloma cell line that does not endogenously produce any immunoglobulin chains), CRL7O3O, COS (e.g., COS1 or COS), PER.C6, VERO, HsS78Bst, HEK-293T, HepG2, SP210, R1.1, BW, LM, BSC1, BSC40, YB / 20, BMT10 and HsS78Bst cells.

[0107] Once the proteins, or antibodies or antigen-binding fragments thereof described herein are produced by recombinant expression, they can be purified by any method known in the art for the purification of proteins or immunoglobulin molecules, such as by chromatography (e.g., ion exchange, affinity, particularly affinity for specific antigens following Protein A, and size exclusion chromatography), centrifugation, differential solubility, or any other standard technique for purifying proteins. Additionally, the proteins, or antibodies or antigen-binding fragments thereof described herein can be fused to heterologous polypeptide sequences described herein or other heterologous polypeptide sequences known in the art to facilitate purification.

[0108] In some embodiments, the antibodies or antigen-binding fragments thereof described herein are isolated or purified. Generally, an isolated protein, or antibody or antigen-binding fragment thereof, is substantially free of other proteins and antibodies or antigen-binding fragments thereof that have antigenic specificities different from those of the isolated antibody or antigen-binding fragment thereof. For example, in some embodiments, preparations of the antibodies or antigen-binding fragments thereof described herein are substantially free of cellular materials and / or chemical precursors.

[0109] 3. Treatment method Treatment of CAH is based on normalizing hormone and steroid levels using various medications, from diagnosis in infancy to adulthood. Glucocorticoids are the current standard of care for CAH and are used both to correct endogenous cortisol deficiency and to reduce elevated ACTH levels in the pituitary gland, which drive increased androgen production. Unlike treatment for Addison's disease (adrenal insufficiency), in which physiologic cortisol replacement is sufficient to reduce ACTH concentrations to normal, treatment for CAH must also reduce ACTH production to control concomitant androgen excess. Thus, the goals of glucocorticoid treatment include cortisol replacement and ACTH suppression to accelerate skeletal maturation and reduce subsequent short stature in both sexes, reduce virilization and menstrual disorders in women, and inhibit the development of testicular adrenal rest tumors in men. Mineralocorticoid replacement is necessary to achieve normal plasma renin activity, which maintains normal blood pressure, electrolyte balance, and blood volume status in patients with the salt-wasting form of CAH.

[0110] Glucocorticoid treatment regimens must support normal physiological function and also ensure that sufficient cortisol is available during events that may elicit a strong stress response (e.g., intercurrent illness, exercise, hypotension). Careful monitoring is also necessary to avoid the development of iatrogenic Cushing's syndrome due to glucocorticoid overtreatment or Addison's syndrome due to undertreatment in an attempt to adequately suppress androgen production.

[0111] Overtreatment with mineralocorticoids can cause hypertension, while undertreatment can lead to hypotension, salt wasting, fatigue, and increased glucocorticoid requirements. Typical laboratory tests to monitor treatment effectiveness include measurement of plasma concentrations of 17-OH, testosterone, free testosterone, androstenedione, 11-oxygenated androgens, renin activity, electrolytes, or a combination of these.

[0112] Adult patients with CAH have a high prevalence of cardiovascular disease risk factors, including obesity, hypertension, and insulin resistance (see, e.g., Kim et al., Semin. Reprod. Med. 27(4):316-21 (2009)). A large cohort study of pediatric and adult CAH patients (n=244) demonstrated that patients were prescribed various glucocorticoid treatment regimens but frequently suffered from poor hormonal control and the aforementioned adverse events (see, e.g., Finkielstain et al., J. Clin. Endocrinol Met. 97(12):4429-38 (2012)).

[0113] Treatment of CAH involves attempting to correct the cortisol deficiency with glucocorticoids (usually hydrocortisone in children, but often more potent agents with narrow therapeutic indices, such as dexamethasone, in adults), and, if needed for salt wasting, mineralocorticoids (usually fludrocortisone). The glucocorticoid doses required to achieve adequate suppression of androgen excess are usually significantly higher than the usual physiologic doses used for cortisol replacement alone in patients with Addison's disease. This increased exposure to glucocorticoids can lead to iatrogenic Cushing's syndrome, increased cardiovascular risk factors, glucose intolerance, and decreased bone mineral density in CAH patients (see, e.g., Elnecave et al., J. Pediatr. Endocrinol. Metab. 21: 1155-62 (2008); King et al., J. Clin. Endocrinol Metab. 91(3): 8656-59 (2006); Migeon et al., Endocrinol. Metab. Clin. North Am. 30: 193-206) (2001)). Recently, best practices for the clinical management of congenital adrenal hyperplasia were published in the Journal of Cliniced Endocrinology and Metabolism (Speiser, PW, et al. J. Clin. Endocrinol Metab. November 2018, 103(11): 1-46). The article is incorporated by reference in its entirety.

[0114] Corticotropin-releasing factor (CRF), also known as corticotropin-releasing hormone (CRH), has been found to produce profound changes in endocrine, nervous, and immune system function. CRF is believed to be the primary physiological regulator of basal and stress-induced release of adrenocorticotropic hormone ("ACTH"), β-endorphin, and other proopiomelanocortin ("POMC")-derived peptides from the prepituitary gland (see, e.g., Vale et al., Science 273: 1394-1397, 1981). Secretion of CRF triggers the release of ACTH from the prepituitary corticotropin gland via binding to the CRFi receptor, a member of the class B family of G protein-coupled receptors.

[0115] The pituitary hormone ACTH, under the control of hypothalamic CRF, stimulates cholesterol uptake and drives the synthesis of pregnenolone, which initiates steroidogenesis in the adrenal gland. The adrenal cortex is composed of three zones, which produce distinct classes of hormones, many of which are driven by ACTH, which mobilizes cholesterol through this pathway. Deficiency of these enzymes, caused by mutation or deletion, results in increased substrate concentrations.

[0116] In the most common form of CAH, caused by mutations or deletions of the 21-hydroxylase gene (CYP21A2), potent androgens are produced by the adrenal glands through the accumulation of steroid precursors, progesterone and 17-hydroxyprogesterone (17-OHP). Plasma levels of 17-OHP can reach 10 to 1,000 times normal levels in these cases. These increases result in the overproduction of androgens, specifically testosterone, free testosterone, androstenedione, 11-oxidized androgens, or a combination of these, causing virilization in women. Additionally, 21-hydroxylase deficiency in CAH leads to insufficient biosynthesis of glucocorticoids and mineralocorticoids, specifically cortisol and aldosterone. Cortisol is an important negative feedback regulator of hypothalamic CRF secretion and pituitary ACTH release. Lack of glucocorticoid synthesis and release eliminates the constraints of the hypothalamus and pituitary, which increases ACTH levels. Excessive ACTH stimulation causes hypertrophy of the zona fasciculata and zona reticularis, resulting in adrenal hyperplasia.

[0117] Patients with CAH are unable to synthesize and secrete normal amounts of glucocorticoids and cortisol due to loss-of-function mutations in CYP21A2. Reduced glucocorticoid levels result in reduced negative feedback to the pituitary gland and brain, causing increased blood ACTH levels, which in turn drives increased adrenal androgen secretion. Increased ACTH levels due to reduced negative feedback of cortisol require CRH, without which cortisol does not rise, as shown in mice with a genetic deficiency of CRH (Muglia et al., Journal of Clinical Investigation, 105(9):1269-1277, 2000). This suggests that blocking CRH action should reduce the increase in blood ACTH levels in CAH patients, resulting in reduced androgen secretion in these patients.

[0118] Additionally, mutations and deletions in the CYP11B1 gene may be associated with CAH. The CYP11B1 gene encodes the enzyme 11-beta-hydroxylase, which is found in the adrenal glands and helps produce cortisol and corticosterone.

[0119] Mutations and deletions of the HSD3B2 gene can be associated with CAH. The HSD3B2 gene encodes the enzyme 3-beta-hydroxysteroid dehydrogenase, which is found in the gonads and adrenal glands and is involved in the production of many hormones, including cortisol, aldosterone, androgens, and estrogens.

[0120] In some embodiments of any of the methods provided herein, CAH is associated with one or more mutations or deletions in a gene selected from the group consisting of CYP21A2, CYP11B1, and HSD3B2. In some embodiments of any of the methods provided herein, CAH is associated with a mutation or deletion in CYP21A2. In some embodiments of any of the methods provided herein, CAH is associated with a mutation or deletion in CYP11B1. In some embodiments of any of the methods provided herein, CAH is associated with a mutation or deletion in HSD3B2.

[0121] The present disclosure relates to a method for treating congenital adrenal hyperplasia (CAH). The method comprises administering to a subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof disclosed herein. In some embodiments, the method comprises administering to a subject a therapeutically effective amount of a pharmaceutical composition of the present disclosure containing the antibody or antigen-binding fragment thereof disclosed herein.

[0122] Provided herein is a method for treating congenital adrenal hyperplasia (CAH), comprising administering an antibody or antigen-binding fragment thereof disclosed herein to normalize or partially normalize the level of a biomarker associated with congenital adrenal hyperplasia. In some embodiments, normalizing or partially normalizing the level of a biomarker comprises reducing the level of an elevated biomarker or increasing the level of a depressed biomarker compared to subjects without CAH.

[0123] Provided herein is a method for treating congenital adrenal hyperplasia in a subject in need thereof, comprising administering an antibody or antigen-binding fragment thereof disclosed herein in an amount sufficient to reduce the level of one or more biomarkers associated with congenital adrenal hyperplasia. In some embodiments, the biomarkers are selected from (a) 17-hydroxyprogesterone (17-OHP), (b) adrenocorticotropic hormone (ACTH), and (c) androstenedione in the subject.

[0124] In some embodiments, the reduction in the level of any of the biomarkers (e.g., any of 17-OHP, ACTH, and androstenedione) is determined by comparing the level of the biomarker measured the day before administration of an antibody or antigen-binding fragment thereof disclosed herein with the level of the biomarker measured the day after administration of an antibody or antigen-binding fragment thereof disclosed herein. The day before administration of an antibody or antigen-binding fragment thereof disclosed herein applies to subjects who have not already been administered an antibody or antigen-binding fragment thereof disclosed herein within at least the past 24 hours.

[0125] In some embodiments of the methods provided herein, the level of 17-hydroxyprogesterone is reduced by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 50%, at least 55%, or at least 60% from the pre-administration level. In some embodiments, the level of 17-hydroxyprogesterone is reduced by at least 25%. In some embodiments, the level of 17-hydroxyprogesterone is reduced by at least 50%. In some embodiments of the methods provided herein, the level of 17-hydroxyprogesterone is reduced by an amount from pre-administration levels of about 10% to about 90%, about 15% to about 90%, about 20% to about 90%, about 25% to about 90%, about 30% to about 90%, about 35% to about 90%, about 40% to about 90%, about 50% to about 90%, about 55% to about 90%, or about 60% to about 90%.

[0126] In some embodiments of the methods provided herein, the level of adrenocorticotropic hormone is reduced by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 50%, at least 55%, or at least 60% from the pre-administration level. In some embodiments, the level of adrenocorticotropic hormone is reduced by at least 25%. In some embodiments, the level of adrenocorticotropic hormone is reduced by at least 40%. In some embodiments, the level of adrenocorticotropic hormone is reduced by at least 50%. In some embodiments of the methods provided herein, the level of adrenocorticotropic hormone is reduced by an amount of about 10% to about 90%, about 15% to about 90%, about 20% to about 90%, about 25% to about 90%, about 30% to about 90%, about 35% to about 90%, about 40% to about 90%, about 50% to about 90%, about 55% to about 90%, or about 60% to about 90% from the pre-administration level.

[0127] In some embodiments, the level of adrenocorticotropic hormone is reduced to a level within the range of adrenocorticotropic hormone expected in a subject without CAH.

[0128] In some embodiments of the methods provided herein, androstenedione levels are reduced by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 50%, at least 55%, or at least 60% from pre-administration levels.In some embodiments, androstenedione levels are reduced by at least 25%.In some embodiments, androstenedione levels are reduced by at least 30%.In some embodiments, androstenedione levels are reduced by at least 50%. In some embodiments of the methods provided herein, androstenedione levels are reduced by an amount of about 10% to about 90%, about 15% to about 90%, about 20% to about 90%, about 25% to about 90%, about 30% to about 90%, about 35% to about 90%, about 40% to about 90%, about 50% to about 90%, about 55% to about 90%, or about 60% to about 90% from pre-administration levels.

[0129] In some embodiments, androstenedione levels are reduced to levels within the range of androstenedione expected in a subject without CAH, ie, below 200 ng / dL.

[0130] Also provided herein is a method of reducing the severity of one or more symptoms selected from hirsutism, precocious puberty, fertility problems, acne, and growth impairment in a subject with classic congenital adrenal hyperplasia, comprising administering an antibody or antigen-binding fragment thereof disclosed herein in an amount sufficient to reduce one or more biomarkers in the subject, e.g., reduce androstenedione in the subject. Growth impairment can refer to, for example, accelerated height velocity, accelerated weight velocity, and / or accelerated bone age.

[0131] Provided herein is a method for reducing the level of one or more biomarkers of CAH for congenital adrenal hyperplasia in a subject with congenital adrenal hyperplasia, comprising administering to the subject an antibody or antigen-binding fragment thereof disclosed herein. In some embodiments, the one or more biomarkers of congenital adrenal hyperplasia are selected from the group consisting of (a) 17-hydroxyprogesterone (17-OHP), (b) adrenocorticotropic hormone (ACTH), and (c) androstenedione.

[0132] Accordingly, there is provided a method of treating congenital adrenal hyperplasia in a subject, comprising: (a) measuring the level of one or more biomarkers selected from (a) 17-hydroxyprogesterone (17-OHP), (b) adrenocorticotropic hormone (ACTH), and (c) androstenedione in a biological sample obtained from the subject; (b) analyzing the level of the one or more biomarkers to determine whether the level of the one or more biomarkers is elevated compared to a healthy subject without congenital adrenal hyperplasia; (c) if the subject is determined to have elevated levels of one or more biomarkers, administering to the subject an antibody or antigen-binding fragment thereof disclosed herein; Provided herein is a method comprising:

[0133] Also provided herein are methods for reducing the dosage of a corticosteroid administered to a subject with congenital adrenal hyperplasia to control the condition, the method comprising administering to the subject an antibody or antigen-binding fragment thereof disclosed herein. In some embodiments, the corticosteroid is a glucocorticoid. In some embodiments, the antibody or antigen-binding fragment thereof and the corticosteroid are administered simultaneously in a single composition in a pharmaceutically acceptable formulation for simultaneous administration, or as separate compositions of the antibody or antigen-binding fragment thereof and the corticosteroid in a pharmaceutically acceptable formulation. In some embodiments, the formulations are administered sequentially in any order.

[0134] Also provided herein is a method for reducing the severity of one or more side effects of glucocorticoid treatment in a subject with congenital adrenal hyperplasia, comprising administering to the subject an antibody or antigen-binding fragment thereof disclosed herein. The long-term effects of glucocorticoid treatment have been well documented in the art (see, for example, Gray, M. et al. (2016): Long-term effect of glucocorticoids, Expert Opinion on Drug Safety. DOI: 10.1517 / 14740338.2016.1140743). Such side effects may affect any biological system, for example, musculoskeletal (e.g., osteoporosis, avascular osteonecrosis, and myopathy), endocrine and metabolic (e.g., hyperglycemia, diabetes, dyslipidemia, weight gain, Cushing's syndrome, Cushingoid features, growth suppression, adrenal suppression), gastrointestinal (e.g., gastritis, peptic ulcer, gastrointestinal bleeding, visceral perforation, fatty liver, pancreatitis), cardiovascular (e.g., hypertension, coronary heart disease, ischemic heart disease, heart failure), dermatological (e.g., dermatoprosis, atrophy of the skin, ecchymoses, purpura, erosions, striae, delayed wound healing, easy bruising, acne, hirsutism, and alopecia), neuropsychiatric (e.g., mood swings, depression, euphoria, mood lability, laability, irritability, akathisia, anxiety, cognitive impairment, psychosis, dementia, and delirium), ophthalmologic (e.g., cataracts, glaucoma, ptosis, mydriasis, opportunistic ocular infections, and central serous chorioretinopathy), and immunologic (e.g., suppression of cell-mediated immunity, predisposition to infection, and reactivation of latent infection).

[0135] Thus, in some embodiments, the side effects of glucocorticoid treatment are selected from osteoporosis, avascular osteonecrosis, myopathy, hyperglycemia, diabetes, dyslipidemia, weight gain, Cushing's syndrome, Cushingoid features, growth suppression, adrenal suppression, gastritis, peptic ulcer, gastrointestinal bleeding, visceral perforation, fatty liver, pancreatitis, hypertension, coronary heart disease, ischemic heart disease, heart failure, skin porousness, atrophy of the skin, ecchymoses, purpura, erosions, striae, delayed wound healing, easy bruising, acne, hirsutism, alopecia, mood swings, depression, euphoria, emotional lability, irritability, akathisia, anxiety, cognitive impairment, psychosis, dementia, delirium, cataracts, glaucoma, ptosis, mydriasis, opportunistic ocular infections, central serous chorioretinopathy, suppression of cell-mediated immunity, predisposition to infection, and reactivation of latent infection, and any combination thereof.

[0136] 4. Glucocorticoids Glucocorticoids are a class of corticosteroids, a class of steroid hormones. Glucocorticoids are corticosteroids that bind to glucocorticoid receptors, which are present in almost all vertebrate cells. In some embodiments, the subject is concurrently receiving glucocorticoids. In some embodiments, the glucocorticoid is selected from cortisol (hydrocortisone), cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, fludrocortisone acetate, and deoxycorticosterone acetate. In some embodiments, the glucocorticoid is cortisol (hydrocortisone). In some embodiments, the glucocorticoid is cortisone. In some embodiments, the glucocorticoid is prednisolone. In some embodiments, the glucocorticoid is dexamethasone.

[0137] In some embodiments, the glucocorticoid dose is measured in hydrocortisone equivalents. In some embodiments, the glucocorticoid dose is measured as a multiple of the upper limit of normal physiological dosing of hydrocortisone equivalents. Any glucocorticoid can be given at a dose that provides approximately the same glucocorticoid effect as normal cortisol production, which is referred to as physiological dosing, replacement dosing, or maintenance dosing.

[0138] In some embodiments, the glucocorticoid dose is a physiological dose measured after a period of administration of the antibody or antigen-binding fragment thereof disclosed herein. In some embodiments, the glucocorticoid dose given to the subject at the same time is a normal physiological dose equivalent to hydrocortisone. In some embodiments, the glucocorticoid dose given to the subject at the same time is determined after a period of administration of the antibody or antigen-binding fragment thereof disclosed herein. In some embodiments, the glucocorticoid dose given to the subject at the same time is determined after a period of administration of the antibody or antigen-binding fragment thereof disclosed herein.

[0139] In some embodiments, the glucocorticoid dose given to the subject concurrently is at the upper limit of the normal physiological dose of hydrocortisone equivalent, hi some embodiments, the glucocorticoid dose given to the subject concurrently is determined after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0140] In some embodiments, the subject exhibits a reduced glucocorticoid load after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the reduced glucocorticoid load is compared to the glucocorticoid load before administration of the antibody or antigen-binding fragment thereof disclosed herein. In some embodiments, one or more symptoms selected from quality of life, fatigue, sleep, insulin resistance, glucose tolerance, glucose control, dyslipidemia, hyperlipidemia, bone mineral density, bone metabolism, fat mass, body weight, central obesity, blood pressure, severe hirsutism, menstrual cycle, control of testicular adrenal rest tumor, and fertility are improved after administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the improvement in the one or more symptoms is compared to the state of the one or more symptoms before administration of the antibody or antigen-binding fragment thereof disclosed herein.

[0141] In some embodiments, the quality of life as measured by EuroQol 5 Dimensions 5 Levels (EQ-5D-5L) in a subject improves after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the improvement in EuroQol 5 Dimensions 5 Levels (EQ-5D-5L) is compared to the EuroQol 5 Dimensions 5 Levels (EQ-5D-5L) result before administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0142] In some embodiments, fatigue is reduced in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the reduction in fatigue is compared to fatigue prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0143] In some embodiments, sleep is increased in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the increase in sleep is compared to sleep before administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0144] In some embodiments, insulin resistance is reduced in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the reduction in insulin resistance is compared to the insulin resistance prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0145] In some embodiments, glucose tolerance is reduced in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the reduction in glucose tolerance is compared to glucose tolerance prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0146] In some embodiments, glucose control is increased in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the increase in glucose control is compared to glucose control prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0147] In some embodiments, lipid levels reflective of dyslipidemia are reduced in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the reduction in lipid levels is compared to lipid levels prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0148] In some embodiments, lipid levels reflective of hyperlipidemia are reduced in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the reduction in lipid levels is compared to lipid levels prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0149] In some embodiments, bone mineral density is increased in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the increase in bone mineral density is compared to bone mineral density prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0150] In some embodiments, bone metabolism is increased in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the increase in bone metabolism is compared to bone metabolism prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0151] In some embodiments, fat mass is reduced in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the reduction in fat mass is compared to the fat mass prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0152] In some embodiments, body weight is reduced in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the reduction in body weight is compared to the body weight prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0153] In some embodiments, central obesity is reduced in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the reduction in central obesity is compared to the central obesity prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0154] A reduced corticosteroid load in growing children can result in increased height growth and increased final adult height.

[0155] In some embodiments, blood pressure is reduced in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the reduction in blood pressure is compared to the blood pressure before administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0156] In some embodiments, severe hypertrichosis is reduced in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the reduction in severe hypertrichosis is compared to the severe hypertrichosis prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0157] In some embodiments, menstrual cycle length is increased in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the increase in menstrual cycle length is compared to menstrual cycle length before administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0158] In some embodiments, control of testicular adrenal rest tumors is increased in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the increased control of testicular adrenal rest tumors is compared to control of testicular adrenal rest tumors prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0159] Administration of antibodies to children may reduce androgen levels, thereby causing a reduction in skeletal maturation and resulting in an increase in final adult height.

[0160] In some embodiments, fertility is increased in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the increase in fertility is compared to fertility prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0161] In some embodiments, gonadotropin levels are increased in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the increase in gonadotropin levels is compared to gonadotropin levels prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0162] In some embodiments, progesterone levels are increased in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the increase in progesterone levels is compared to the progesterone levels prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0163] In some embodiments, semen levels are increased in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the increase in semen levels is compared to semen levels prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0164] In some embodiments, LH (progesterone) levels are increased in a subject after a period of administration of an antibody or antigen-binding fragment thereof disclosed herein, wherein the increase in LH levels is compared to the LH levels prior to administration of an antibody or antigen-binding fragment thereof disclosed herein.

[0165] In some embodiments, the subject is an adult. In some embodiments, the subject is over 18 years old. In some embodiments, the subject is a female. In some embodiments, the subject is a male. In some embodiments, the subject is an adolescent under 20 years old or a child under 13 years old.

[0166] 5. Detection Method It is sometimes desirable to detect the presence of CRH in a sample or to measure the amount of CRH. In this regard, the present disclosure provides a method for measuring the amount of CRH in a sample using the antibodies or antigen-binding fragments thereof disclosed herein. To determine the measured amount of CRH, a biological sample from a mammalian subject is contacted with an anti-CRH antibody (or antigen-binding fragment thereof) described herein for a time sufficient to allow the formation of an immune complex. The immune complex formed between the antibody and CRH in the sample is then detected. The amount of CRH in a biological sample may be quantified by measuring the amount of immune complex formed between the antibody and CRH. For example, the antibody can be quantitatively measured if it has a detectable label, or a secondary antibody can be used to quantify the immune complex.

[0167] In some embodiments, the biological sample comprises a tissue sample, a cell sample, or a bodily fluid sample, such as blood, saliva, serum, or plasma.

[0168] The conditions for incubating the antibody with the test sample vary. Incubation conditions depend on the format used in the assay, the detection method used, and the type and nature of the antibody used in the assay. Those skilled in the art will recognize that any one of the commonly available immunoassay formats can be easily adapted to use the antibody (or its fragment) of the present disclosure. Examples of such assays can be found in Chard, T., An Introduction to Radioimmunoassay and Related Techniques, Elsevier Science Publishers, Amsterdam, The Netherlands (1986); Bullock, GR et al., Techniques in Immunocytochemistry, Academic Press, Orlando, FL Vol.1 (1982), Vol.2 (1983), Vol.3 (1985); Tijssen, P., Practice and Theory of Immunoassays: Laboratory Techniques in Biochemistry and Molecular Biology, Elsevier Science Publishers, Amsterdam, The Netherlands (1985). The test sample used in the above-described methods will vary based on the assay format, nature of the detection method, and the tissues, cells, or body fluids used as the sample to be assayed.

[0169] The assays described herein may be useful, for example, in assessing the effectiveness of a particular therapeutic treatment regime in animal studies, clinical trials, or in monitoring the treatment of an individual patient.

[0170] In some embodiments, the CRH or anti-CRH antibody (or antigen-binding fragment thereof) is attached to a solid support, and binding is detected by detecting a complex of CRH and the antibody (or antigen-binding fragment thereof) on the solid support. The antibody (or antigen-binding fragment thereof) may contain a detectable label, and binding is detected by detecting the label of the CRH-antibody complex.

[0171] The detection of the presence or absence of CRH-antibody complexes can be achieved using any method known in the art. For example, the transcripts obtained by reporter gene transcription assay of CRH peptides interacting with target molecules (e.g., antibodies) typically encode directly or indirectly detectable products (e.g., β-galactosidase activity and luciferase activity). In cell-free binding assays, one of the components usually contains or is bound to a detectable label. A wide variety of labels can be used, including those that provide direct detection (e.g., radioactivity, luminescence, optical or electron density) or indirect detection (e.g., epitope tags, e.g., FLAG epitopes, enzyme tags, e.g., horseradish peroxidase). The label can be bound to the antibody or incorporated into the antibody structure.

[0172] Depending on the nature of the label and other assay components, various methods can be used to detect the label.For example, the label can be detected while bound to the solid substrate or after being separated from the solid substrate.The label can be detected directly through optical or electron density, radioactive emission, non-radioactive energy transfer, or indirectly through antibody conjugates or streptavidin-biotin conjugates.Methods for detecting labels are well known in the art.

[0173] 6. Kit Once the pharmaceutical composition is formulated, it can be stored in a sterile vial as a solution, suspension, gel, emulsion, solid, crystalline, or dehydrated or lyophilized powder. Such formulations can be stored either in a ready-to-use form or in a form (e.g., lyophilized) that is reconstituted before administration. The present invention also provides kits that produce single-dose administration units. The kits of the present disclosure can contain a first container with a dried protein and a second container with an aqueous formulation. In certain embodiments, kits containing single-chamber and multi-chamber pre-filled syringes (e.g., liquid syringes and lyosyringes) are provided. [Example]

[0174] The following examples are provided to further illustrate aspects of the present disclosure and are not intended to bind the present disclosure to any particular application or theory of operation.

[0175] [Example 1] Generation of CRH knockout mice on the H2L2 Harbour Mice® background Corticotropin-releasing hormone (CRH) knockout (KO) mice were generated on the H2L2 Harbour Mice® background using standard transgenic methods described in "Manipulating the Mouse Embryo," a laboratory manual by Nagy et al., Cold Spring Harbor Laboratory Press. Generation of CRH KO H2L2 mice was performed under animal protocol 16-512-11 approved by the Dutch Central Authority for Scientific Procedures on Animals of (CCD) and the EDC animal facility under animal license ADV101002016512.

[0176] Briefly, superovulated H2L2 Harbour Mice® females and males were used for mating. The collected fertilized eggs were used for microinjection of 20 ng / microliter of two guide RNAs (gRNAs): gRNA1 Mm.Cas9.CRH.1.AA and gRNA2 Mm.Cas9.CRH.1.AB, and CAS9 protein in a microinjection buffer of 5 mM tris(hydroxymethyl)aminomethane (TRIS), 0.1 mM ethylenediaminetetraacetic acid (EDTA) in ultrapure water adjusted to pH 7.4. The following gRNA1 sequence was used: gRNA1=Sequence-Mm.Cas9CHR.1.AA: 5'-AltR1 / rArArC rUrCrC rArCrG rCrCrC rCrUrC rArCrC rGrCrG rUrUrU rUrArG rArGrC rUrArU rGrCrU / altR2 / -3' gRNA2=Seq-Mm.Cas9.CRH.1.AB: 5'-AltR1 / rUrCrA rCrCrC rArUrG rCrGrG rArUrC rArGrArArCrG rUrUrU rUrArG rArGrC rUrArU rGrCrU / altR2 / -3'

[0177] Guide RNA and CAS9 were obtained from Integrated DNA Technologies (IDT).

[0178] Injected eggs were transferred to foster mothers (B6CBAF1). Knockout offspring were confirmed from toe DNA by amplification of DNA spanning the gRNA region. Polymerase chain reaction (PCR) fragments smaller than the predicted size indicated the deletion in the founder mice. PCR primers CRHbp70F and CRHbp215R, which flank gRNA1, were used to amplify the predicted size of 146 bp from wild-type H2L2 Harbour Mice® (Figure 2A, left panel). PCR primers CRHbp208F and CRHbp310R were used to amplify the predicted size of 103 bp from wild-type H2L2 Harbour Mice® (Figure 2A, center panel). PCR primers CRHbp70F and CRHbp310R resulted in a PCR fragment size of 241 bp (Figure 2A, right panel). PCR products smaller than the predicted size indicated mutations. Founder mice 1, 2, 4, and 5 were confirmed to carry the mutation. PCR products that deviated from the predicted size of the wild-type were further analyzed by cutting the PCR amplification product from an agarose gel, isolating the DNA, and sequencing using the same primers used for amplification. Founder mouse 5 from litter 12198 in Figure 2B has an 11-bp deletion (5'-CAGCCGGTTCT-3') (SEQ ID NO: 465) and was used for subsequent breeding.

[0179] Rescue of CRH knockout H2L2 Harbour Mice® Rescue and immunization of CRH knockout mice were performed under Animal License ADV101002016512, Animal Treatment Protocol 15-512-19. Briefly, homozygous CRH knockout H2L2 Harbour Mice® males and females were housed in a 1:1 cage. Upon observation of a mating plug (0.5 days after conception), the females were removed from the cage and separated. After 10 days, cortisol (30 μg / ml) was provided ad libitum to pregnant females in their drinking water (dark bottles). This resulted in the birth of live knockout offspring, which were used for immunization.

[0180] Immunization of H2L2 Harbour Mice® with CHR knockout Ten homozygous CRH knockout H2L2 Harbour Mice® of both sexes, approximately 8 weeks of age, were immunized on days 0, 14, 28, 42, and 50 by subcutaneous (SC) and intraperitoneal (IP) injection of a maximum volume of 100 μl of CRH-KLH conjugate.

[0181] The following conjugates were used: CRH-KLH: SEEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRKLMEII-KLH (C-terminus) (GenScript) (SEQ ID NO: 467)

[0182] The CRH-KLH conjugate was mixed with the adjuvant Stimmune for the first injection and with the adjuvant Ribi for subsequent boosts. The final boost was IP. Blood samples were collected after the fourth injection and at the end of the serial immunization experiment. Three to five days after the final IP injection, mice were euthanized, and their spleens and lymph nodes were used for hybridoma production in fusion experiments with myeloma fusion partners according to standard protocols. Serum was screened for anti-CRH antibody titers and later for hybridomas by enzyme-linked immunosorbent assay (ELISA) using biotinylated CRH captured on streptavidin (SA)-coated plates. The detailed protocol is as follows:

[0183] CRH ELISA ELISA plates were coated with 5 μg / ml streptavidin in phosphate-buffered saline (PBS) (Sigma 189730-1mg) overnight at 4°C or for 2 hours at room temperature (RT). 50 μl / well was used for 96-well plates, and less for 384-well plates. After incubation, the streptavidin solution was removed and biotinylated peptide was added (ThermoFisher; biotin is at the N-terminus of the peptide). After a 30-minute incubation at RT, the plates were washed with 1x phosphate-buffered saline (PBS) / 0.1% Tween-20 and blocked with 1% milk / 1% bovine serum albumin (BSA) in the wash solution. After washing with 3x PBS / 0.1% Tween-20, 50 μl of antibody was added in blocking buffer and incubated for 2 hours at RT. Plates were then washed with 3x PBS / 0.1% Tween-20, and 50 μl of anti-rat IgG-HRP (horseradish peroxidase) was added at a 1:2,000 dilution and incubated for 2 h at RT. After washing with 3x PBS / 0.1% Tween-20, 50 μl of peroxidase substrate (POD or equivalent) was added, and absorbance was measured at 450 nm.

[0184] Immunization of Balb / c wild-type mice Balb / c wild-type mice were immunized with CRH-KLH protein to generate anti-CRH antibodies. CRH and KLH conjugates were N-terminally conjugated CRH (peptide 1: KLH-CRH) or C-terminally conjugated CRH (peptide 2: CRH-KLH) via the extra-N- or C-terminal cysteines (C) of CRH. 50 μg of protein was administered to each mouse for the initial challenge IP in Freund's complete adjuvant (CFA, Sigma, catalog no. F5881), followed by 25 μg IP in Ribi (Sigma S6322) for the subsequent boost. Boosts were performed four times every two weeks. Mouse sera were sampled at the indicated time points and titrated by indirect ELISA to test for binding to human CRH.

[0185] Mice with high serum titers and specific immune responses to human CRH were selected and boosted with 25 μg of protein in PBS. Three days later, the mice were sacrificed and spleen cells and lymph nodes were collected.

[0186] Preparation of CRH-KLH conjugate KLH carrier protein (Sigma, H7017-50MG) was weighed and dialyzed against 0.1 M sodium phosphate buffer, pH 7.5. Sulfo-MBS (BBI, C100314-0050) solution was added to the KLH carrier protein solution at a 1:10 ratio. After 1 h of incubation at room temperature, the reaction mixture was applied to a G-50 desalting column (Sigma, G50150-10G) pre-equilibrated with conjugation buffer (0.1 M sodium phosphate buffer, pH 6.5, with 1 mM EDTA). Fractions corresponding to the first protein peak based on A280 nm were collected. KLH carrier protein-MBS and CRH peptides bearing an extra cysteine ​​at either the N- or C-terminus were mixed and incubated at room temperature for 2-4 h. The reaction was terminated by the addition of 2-mercaptoethanol (Sigma, M3148-25ML) to a final concentration of 10 mM and incubated at room temperature for >30 min. The CRH-KLH conjugate was purified using a Sephadex G-25 desalting column to separate the unconjugated peptide from the CRH-KLHJ conjugate. The fractions corresponding to the first protein peak were collected. The protein concentration of the CRH-KLH conjugate was determined using a NanoPhotometer.

[0187] The following peptides were used for immunization of Balb / c wild-type mice: Peptide 1 (KLH-CRH) (SEQ ID NO: 468): KLH-C-SEEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRKLMEII-CONH2 Peptide 2 (CRH-KLH) (SEQ ID NO: 469): SEEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRKLMEII-C-KLH

[0188] The following peptides were used in the ELISA: Peptide 3 (Biotin-CRH) (SEQ ID NO: 470): Biotin-K-SEEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRKLMEII-CONH2 Peptide 4 (CRH-biotin) (SEQ ID NO: 471): SEEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRKLMEII-K-Biotin Biotin-CRH (SEQ ID NO: 472): Biotin-SEEPPISLDLTFHLLREVLEMARAEQLAQQAHSNRKLMEII-COOH (ThermoFisher)

[0189] Single B cell screening based on the Beacon® Optofluidic System We used a nanofluidic optoelectronic B lymphocyte antibody screening technology (NanOBlast) built around the commercially available Beacon™ (Berkeley Lights) and integrated culture and imaging platform.

[0190] The NanOBlast workflow begins with the generation of antigen-specific antibody-secreting cells (ASCs) via in vivo immunization. Selectively enriched, antigen-experienced murine ASCs were harvested from the spleen and lymph nodes. ASCs were microfluidically transferred into the chip and isolated in individual nanopens for screening via OptoElectroPositioning (OEP). Antigen-specific IgG-secreting ASCs were detected using a bead-based fluorescent color binding assay, resulting in a characteristic fluorescent bloom. Individual cells of interest were then removed from the chip using OEP and directly exported into a 96-well plate containing cell lysis buffer. Antibody heavy chain variable domain (VH) and light chain variable domain (VL) sequences were recovered using single-cell rapid amplification of cDNA ends (RACE), cloned, and recombinantly expressed as standard antibodies using standard methods. The recombinant antibodies were used for binding confirmation and final validation in relevant downstream assays.

[0191] Antibody production and purification Recombinant plasmids encoding target antibodies were transiently co-transfected into HEK293-6E or ExpiCHO cell cultures using polyethyleneimine (PEI) (Polyscience, 24885). One day before transfection, cells were seeded into Corning Erlenmeyer flasks. On the day of transfection, recombinant plasmids encoding target proteins and transfection reagents were mixed at an optimal ratio (heavy chain:light chain = 2:3) and then added to the seed flasks for transfection. Cell culture supernatants were collected on day 6 and used for purification. The cell culture broth was centrifuged and subsequently filtered. The filtered cell culture supernatant was loaded onto an affinity purification column. After washing and elution, the eluted fractions were pooled and buffer exchanged with storage buffer. The purified proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and size-exclusion chromatography-high-performance liquid chromatography (SEC-HPLC) analysis to determine molecular weight and purity. The concentration was determined by absorbance at 280 nm.

[0192] Antibody modification The heavy chain variable domain (VH) and light chain variable domain (VL) sequences of the anti-CRH chimeric antibody were further optimized by humanization and post-translational modification (PTM) removal procedures. In the humanization procedure, the VH and VL sequences were first aligned to the closest human germline sequences using an algorithm, such as NCBI / Ig-BLAST, and then positions within the framework regions were reverted to the human germline sequences. All major backmutation positions were scanned using Chothia numbering. Mismatches were replaced with mouse counterpart residues. Antibodies composed of the humanized sequence variants were then recombinantly produced using standard molecular biology techniques.

[0193] For PTM removal, the VH and VL sequences were scanned for the presence of PTM motifs, such as isomerization motifs (e.g., DG). "Hot spot" residues (e.g., D or G in the DG motif) were mutated to either the counterpart residue in the germline sequence or another residue with similar biophysical properties. Antibodies containing the post-PTM removal sequence variants were then recombinantly produced using standard molecular biology techniques.

[0194] Binding of anti-CRH antibodies to CRH or human UCN1 by ELISA Streptavidin (Thermo, catalog: 21125) was diluted in PBS to a concentration of 2 μg / ml. 100 μl of diluted streptavidin was added per well to an ELISA microplate, and the plate was incubated overnight at 4°C. The plate was blocked with ELISA blocking solution (containing 2% w / v BSA, 0.05% (v / v) Tween-20, pH 7.4 PBS buffer) at 37°C for 1 hour, and then incubated with 0.5 μg / mL CRH (NT biotin), 0.5 μg / mL CRH (CT biotin), 1 μg / mL human UCN1 (NT biotin), or 1 μg / mL human UCN1 (CT biotin) for 1 hour at 37°C. The plate was then washed and incubated with 15 μg / ml (100 nM, 10-fold dilution, 8-point) diluted anti-CRH antibody (Ab) at 37°C for 1 hour. Subsequently, the plate was washed and incubated with HRP-conjugated goat anti-human IgG (H+L) antibody (Jacson, catalog: 109-035-088) at 37°C for 1 hour. 100 μL of 3,3',5,5'-tetramethylbenzidine (TMB) substrate (Biopanda, catalog: TMB-S-003) was added, and the plate was incubated at room temperature for 15 minutes. The reaction was terminated by adding 100 μL of ELISA stop solution (Solarbio, catalog: C1058), and the optical density at 450 nm (OD450nm) was determined using an ELISA plate reader (Molecular Devices, Spectra max384plus).

[0195] Plasmid and cell line preparation The CHO-K1 (Cat. No. CCL-61) cell line was obtained from the American Type Culture Collection (ATCC). pGL4.29 / luc2P / CRE / Hygro (Cat. No. E8471) and Bio-Glo Luciferase Assay System (Cat. No. G7940) were obtained from Promega. Plasmids pcDNA3.1-human CRHR1 (clone ID: OHu27188C), pcDNA3.1-human CRHR2 (clone ID: OHu10803), pcDNA3.1-mouse CRHR1 (clone ID: Omu00753), and pcDNA3.1-mouse CRHR2 (clone ID: Omu23246) were synthesized using GenScript® and are G418-resistant. Human CRH peptide was obtained from Peptide International (Cat. No. 4136-s). Forskolin (catalog number 1099) and a human CRHR1 antagonist small molecule (antalarmin, catalog number 2778) were obtained from Tocris. Anti-human CRHR1 antibody (catalog number MAB3930) was obtained from R&D System. Electroporation equipment (Elector Cell Manipulator, catalog number ECM630) and electroporation cuvettes plus (2 mm gap cuvettes, catalog number 45-0125) were used from BTX. Electroporation buffer (Ingenio Solution, catalog number MIR50111) was obtained from Mirus, and sheep anti-human CRH polyclonal antibody was obtained from Salk Institute.

[0196] Generation of CHO-CRE luciferase human CRHR1, human CRHR2, mouse CRHR1, and mouse CRHR2 reporter cell lines To generate CHO-CRE luciferase reporter stable cell lines, a mixture of 2 million CHO-K1 cells and 10 μg of pGL4.29 / luc2P / CRE / Hygro plasmid in 100 μL of Ingenio solution was transferred to an electroporation cuvette and given one or two pulses using an electroporator (950 μF and 120 V). The treated cells were transferred to a 6-well plate in 2 mL of Dulbecco's Modified Eagle Medium (DMEM) / F12 with 10% fetal bovine serum (FBS). After 24 hours, 500 μg / mL of the selection antibody hygromycin was added to the cells. This selection took approximately 10–14 days. Using matrix dilution, pools of CHO-CRE cells were subcloned into 96-well plates. When selected colonies became evident, individual stable clones were picked. Positive clones were verified using a dilution series of forskolin. Verified positive clones were frozen for future use.

[0197] To generate the CHO-CRE-hCRHR1, -hCRHR2, -mCRHR1, and -mCRHR2 reporter cell lines, pcDNA3.1-hCRHR1, -hCRHR2, -mCRHR1, and -mCRHR2 were introduced into the CHO-CRE cells established above by electroporation as described above. Selection antibodies, hygromycin at 500 μg / mL and G418 at 1,000 μg / mL, were used. Each established stable cell line was characterized by both CRH and forskolin stimulation.

[0198] Development of luciferase reporter assays Forskolin stimulates adenylate cyclase, increasing intracellular cyclic adenosine monophosphate (cAMP) levels in cells. To verify whether cells respond to elevated cAMP, CHO-CRE cells were stimulated with a range of concentrations of forskolin, and cAMP-dependent luciferase expression was determined using the Bio-Glo Luciferase Assay System. CHO-CRE cells increased luciferase expression in a dose-dependent manner in response to forskolin stimulation (data not shown). CRH binds to CRHR1 and CRHR2, which are Gαs-coupled G protein-coupled receptors (GPCRs).

[0199] Activated GPCR proteins increase intracellular cAMP production. CHO-CRE-hCRHR1 stable clone 11 was generated by transfecting pcDNA3.1-human CRHR1 under G418 and hygromycin selection. Human CRHR1 expression in CHO-CRE-hCRHR1 clone 11 was confirmed by flow cytometry (data not shown). To further verify that the cell line was functional, CHO-CRE-hCRHR1 clone 11 cells were treated with forskolin and CRH peptide. CHO-CRE-hCRHR1 clone 11 cells showed a dramatic increase in luciferase expression upon CRH treatment compared to the parent CHO-CRE, while both cell lines showed comparable levels of intracellular cAMP in response to forskolin treatment. Subsequently, CHO-CRE-hCRHR2 clone 32, -mCRHR1 clone 22, and -mCRHR2 clone 8 cell lines were established using similar methods and validated by CRH treatment (data not shown).

[0200] Blocking CRHR1 with small molecules or neutralizing CRH peptides can reduce luciferase expression when the CRH-CRHR1 system is activated in CHO-CRE-hCRHR1 cells. The small molecule CRHR1 antagonist antalarmin blocked CRH-mediated luciferase expression in CHO-CRE-hCRHR1 clone 11 (data not shown). Furthermore, sheep anti-CRH polyclonal antibodies reduced CRH-mediated luciferase expression in these cells (data not shown). Similarly, sheep anti-CRH polyclonal antibodies blocked CRH-mediated luciferase expression in CHO-CRE-hCRHR2, CHO-CRE-mCRHR1, and CHO-CRE-mCRHR2 cells (data not shown).

[0201] Affinity determination by Octet Octet Red 96e was used for affinity measurements. Octet running buffer was prepared by diluting kinetic buffer 10X (Fortebio, catalog no. 18-1105) in PBS according to the manufacturer's instructions. Two columns of SA sensors (Fortebio, catalog no. 18-5019) were hydrated in running buffer for 10 minutes. One column was for ligand capture, and the other was for reference. Antibodies were diluted 2-fold and added to a 96-well plate. 30 nM biotinylated CRH (GL Biochem, catalog no. 784751) was captured by the first column of the SA sensors for 20 seconds, while the reference sensor was immersed in the buffer well. After a 60-second baseline step, the sensors were immersed in the analyte, followed by the buffer well. Association and dissociation times were 180 and 800 seconds, respectively. The sensors were regenerated with glycine pH 1.5 (Cytiva, catalog no. BR-1003-54) before the next cycle. Equilibrium dissociation constant (KD) values ​​were evaluated using Data Analysis Software 11.0 and a fitting model of 1:1 global fitting. The signals of the reference well and reference sensor were subtracted by selecting "double referencing" before data fitting.

[0202] Blocking activity of CRH-induced signaling in reporter cell lines (CHO-CRE-hCRHR1, CHO-CRE-hCRHR2, CHO-CRE-mCRHR1, and CHO-CRE-mCRHR2) Cells were digested with trypsin-EDTA (Life Technologies, Catalog No. 12604-013), centrifuged at 1,000 rpm for 5 minutes, resuspended in growth medium, counted using a cell counter, and seeded into a 96-well plate at a concentration of 5,000 cells per 100 μl and incubated overnight. 50 μl of 4× serially diluted antibody was added to the plate containing CHO-CRE-hCRHR1 cells (8-point, 1 / 4 dilution with growth medium), and 50 μl of 4×CRH (4× is 2 nM, final 0.5 nM) was added. Similarly, 4×CRH (4× is 0.4 nM, final 0.1 nM) was added to CHO-CRE-hCRHR2 cells and CHO-CRE-mCRHR1 cells, and 4×CRH (4× is 20 nM, final 5 nM) was added to CHO-CRE-mCRHR2 cells. The plate was then incubated at 37°C in a CO2 incubator for 4-6 hours. 60 μl of culture medium was removed and 60 μl of Bio-Glo luciferase was added, followed by incubation at RT for 10 minutes. The plate was read using a luciferase plate reader (EnVison, PerkinElmer).

[0203] ELISA for human UCN1, human UCN2, and human UCN3 Streptavidin (Thermo, catalog: 21125) was diluted in PBS to a concentration of 2 μg / mL. 100 μL of diluted streptavidin was added per well to an ELISA microplate, and the plate was incubated overnight at 4°C. The plate was blocked with ELISA blocking solution (containing 2% w / v BSA, 0.05% (v / v) Tween-20, pH 7.4 PBS buffer) at 37°C for 1 hour and then incubated with 1 μg / mL hUCN1(NT biotin), hUCN1(CT biotin), hUCN2(NT biotin), hUCN2(CT biotin), hUCN3(NT biotin), or hUCN3(CT biotin) for 1 hour at 37°C. The plate was then washed and incubated with 100 nM and 10 nM anti-CRH antibody at 37°C for 1 hour. The plate was then washed and incubated with HRP-conjugated goat anti-human IgG (H+L) antibody (Jacson, catalog: 109-035-088) for 1 hour at 37°C. 100 μL of TMB substrate (Biopanda, catalog: TMB-S-003) was added, and the plate was incubated for 15 minutes at room temperature. 100 μL of ELISA stop solution (Solarbio, catalog: C1058) was added to terminate the reaction, and the OD450nm was determined using an ELISA plate reader (Molecular Devices, Spectra max384plus).

[0204] Human UCN1-induced luciferase reporter assay CHO-CRE-hCRHR1 and CHO-CRE-hCRHR2 cells were harvested with trypsin-EDTA (Life Technologies, catalog number: 12604-013). Cells were centrifuged at 1,000 rpm for 5 minutes and resuspended in growth medium. Cells were counted using a cell counter and seeded at 5,000 cells per 100 μl into a 96-well plate and incubated overnight. 50 μl of 4× serially diluted antibody (8-point, 1 / 3 dilution with growth medium) or Cp376395 hydrochloride (TOCRIS, catalog number: 3232) (8-point, 1 / 10 dilution with growth medium) was added to the plate, followed by the addition of 50 μl of 4× hUCN1 (4× is 4 nM, final 1 nM) to CHO-CRE-hCRHR1 or CHO-CRE-hCRHR2 cells. The plates were then incubated at 37°C in a CO2 incubator for 4-6 hours. Following this, 60 μl of culture medium was removed and 60 μl of Bio-Glo luciferase was added, followed by incubation at RT for 10 minutes. The plates were then read on a luciferase plate reader (EnVison, PerkinElmer).

[0205] In vivo efficacy model—wild-type female C57BL / 6 mice and Mrap1KO mice Eight- to ten-week-old wild-type female C57BL / 6 mice were obtained from Charles River Laboratories. Mrap1KO mice were obtained from Queen Mary University of London, UK. The small molecule CRHR1 antagonist, SSR125543A, was obtained from Axon Medchem (catalog number Axon1799). Mouse corticosterone (55-Corms-E01) and adrenocorticotropic hormone (ACTH) ELISA kits (21-ACTHU-E01) were obtained from ALPCO, USA. All procedures were performed in accordance with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The protocol was approved by the Charles River Laboratories Institutional Animal Care and Use Committee (IACUC).

[0206] Breeding, ear tagging, and genotyping of Mrap1KO mice To generate Mrap1KO mice, Mrap1- / - male mice were bred with Mrap1+ / - female mice. When female mice became pregnant and pregnancy was palpable (approximately embryonic day 10), corticosterone hormone replacement was administered to the breeding cages by administering 50 μg / ml of drinking water. Drinking water was replaced weekly, and treatment was stopped at weaning. Mice were uniquely identified by ear tags at the time of tail clipping to match the future genotype of each mouse. DNA was extracted from tail tissue and used for genotyping the mice.

[0207] Genotyping of Mrap1− / − mice was performed using PCR and the following PCR primers: Wild-type allele of exon 1 of mrap1: Forward 5'-GCGTCTCTTAGTAGCCTTTGG (SEQ ID NO: 473) Reverse 5'-CTTGTTGGCTTTCAGCTTCT (SEQ ID NO: 474) Mutant allele of Mrap1: Forward 5'-GAACTTGCCTGGTCAACTGTTAAAAGGAC (SEQ ID NO: 475) Reverse 5'-TCTTTAGGCTCACTTCCTCCACTGACC (SEQ ID NO: 476) PCR band sizes: wild type allele is 345 bp, mutant allele is 459 bp.

[0208] Physical restraint of wild-type mice To determine the effect of painless physical restraint on stress responses, mice were restrained in a 50 ml polypropylene ventilation tube with the end cut with a razor blade, allowing the mouse to breathe easily for 15–20 min. The mouse was directed toward the open tube and allowed to crawl through. A piece of paper towel was placed behind the mouse to prevent it from returning and the tube was tightened with a cap. The mouse was returned to its cage and placed ventral-side down on the floor. This stressor was not considered painful because the mouse was confined to a confined space (the ventilation restraint tube) but not in an unnatural physical position. Researchers were present in the room for all procedures. At the end of the procedure, the mouse was removed by its tail.

[0209] Effect of CRH Ab on wild-type mice in a restraint stress model On day 1, each mouse was intraperitoneally injected with 20 mg / kg of anti-CRH mAb, hIgG1, or PBS (five mice per group). On days 2 or 3 and 14, each mouse was subjected to the physical restraint stress described above for 15 or 20 minutes. Immediately after restraint stress, 200 μl of blood was collected from the submandibular vein of each mouse into EDTA tubes. Plasma was separated using refrigerated centrifugation (500 × g at 4 °C for 10 minutes) and flash-frozen on dry ice for subsequent ACTH and corticosterone ELISA measurements.

[0210] Effect of anti-CRH Ab in the Mrap1KO mouse model On day 1, each mouse was injected (IP) with 20 mg / kg of anti-CRH Ab or hIgG1 (5 or 6 mice per group). On days 2 or 3, 14, and 28, 200 μl of blood was collected from the submandibular vein of each mouse into EDTA tubes. Plasma was separated using refrigerated centrifugation (500 × g for 10 min at 4 °C) and flash-frozen on dry ice for subsequent ACTH ELISA measurements.

[0211] Side-by-side comparison of anti-CRH mAb and SSR125543A in a wild-type mouse restraint stress model On day 1, half of the mice were injected IP with 20 mg / kg anti-CRH Ab or hIgG1 (five mice per group). On day 3, the remaining half of the mice were injected IP with 30 mg / kg SSR125543A or vehicle (five mice per group). Two hours later, the four groups of mice were subjected to restraint stress as described above for 20 minutes. Immediately after restraint stress, 200 μl of blood was collected into EDTA tubes from the submandibular vein of each mouse in all four groups. Plasma was separated as described above and flash-frozen on dry ice for subsequent ACTH and corticosterone ELISA measurements. On day 4, SSR125543A- or vehicle-treated mice were subjected to a second restraint stress for 20 minutes, and plasma was collected. Two weeks later, anti-CRH mAb- or hIgG1-treated mice were subjected to physical stress for 20 minutes, followed by plasma collection.

[0212] In vivo PK studies Dosing and Blood Collection For each antibody tested, six female C57BL / 6 mice weighing 18-22 grams were selected and administered a single dose of 5 mg / kg intravenously. Whole blood was collected from three mice before administration, 15 minutes, 24 hours (day 1), 4 days, and 10 days after administration. Whole blood from the remaining three mice was collected before administration, 5 hours, 2, 7, and 14 days after administration. The whole blood was allowed to clot for 30 minutes and then centrifuged. The separated serum was then frozen at -80°C until analysis.

[0213] Analysis method Drug concentrations in mouse serum were quantitatively determined using ELISA. Antibodies (with human Fc) tested in mouse serum were captured with a goat anti-human Fc polyclonal antibody (Rockland, no. 609-101-017) pre-coated on a 96-well plate. An HRP-labeled goat anti-human (H+L) secondary antibody (Abcam, no. ab97175) was then added to the wells. The plate was then sealed and incubated at 37°C for 30 minutes, followed by washing and the addition of TMB substrate and ELISA stop solution. The 96-well plate was read at wavelengths of 450 / 630 nm using a SpectraMax M2. Data were processed using Soft Max Pro.

[0214] Pharmacokinetic (PK) parameters were analyzed by non-compartmental model (NCA) Phoenix WinNonlin version 8.2.

[0215] Determination of CRH binding domain of anti-CRH antibody by ELISA Streptavidin (Thermo, catalog: 21125) was diluted in PBS to a concentration of 2 μg / ml. 100 μl of diluted streptavidin was added per well to an ELISA microplate, and the plate was incubated overnight at 4°C. The plate was blocked with ELISA blocking solution (containing 2% w / v BSA, 0.05% (v / v) Tween-20, pH 7.4 PBS buffer) for 1 hour at 37°C and then incubated with 1 μg / mL of the N-terminal region plus central region (CT biotin), the central region only (NT biotin or CT biotin), or the central region plus C-terminal region (NT biotin) of CRH for 1 hour at 37°C. The plate was then washed and incubated with 15 μg / ml (100 nM, 10 dilutions, 8 points) diluted anti-CRH antibody (Ab) for 1 hour at 37°C. The plate was then washed and incubated with HRP-conjugated goat anti-human IgG (H+L) antibody (Jacson, catalog number: 109-035-088) for 1 hour at 37°C. 100 μL of 3,3',5,5'-tetramethylbenzidine (TMB) substrate (Biopanda, catalog number: TMB-S-003) was added, and the plate was incubated for 15 minutes at room temperature. The reaction was terminated by adding 100 μL of ELISA stop solution (Solarbio, catalog number: C1058), and the optical density at 450 nm (OD450nm) was determined using an ELISA plate reader (Molecular Devices, Spectra max384plus).

[0216] Epitope binding by competitive ELISA To confirm epitope binding, a competitive ELISA was performed. Specifically, 100 μl / well of 1 μg / ml capture antibody was coated onto a 96-well plate overnight at 4°C. The plate was blocked with ELISA blocking solution (containing 2% w / v BSA, 0.05% (v / v) Tween-20, pH 7.4 PBS buffer) at 37°C for 1 hour. Next, 50 μl / well of a mixture of competitive antibody (40 μg / mL) and 50 μl / well of biotin-labeled CRH at the appropriate concentration was added to the 96-well plate, followed by incubation at 37°C for 1 hour. The plate was then washed and incubated with streptavidin peroxidase (SIGMA, catalog: S2438-250UG) at 37°C for 1 hour. 100 μL of 3,3',5,5'-tetramethylbenzidine (TMB) substrate (Biopanda, catalog: TMB-S-003) was added, and the plate was incubated for 15 minutes at room temperature. The reaction was terminated by adding 100 μL of ELISA stop solution (Solarbio, catalog: C1058), and the optical density at 450 nm (OD450nm) was determined using an ELISA plate reader (Molecular Devices, Spectra max384plus).

[0217] Epitope binning using Octet Epitope binding was performed with Octet Red 96e. Running buffer was prepared by diluting kinetic buffer 10x (Fortebio, catalogue no. 18-1105) in PBS according to the manufacturer's instructions.

[0218] The SA sensor (Fortebio, catalog no. 18-5019) was hydrated with running buffer for 10 minutes. The in-tandem epitope binding assay involved a four-step binding cycle: 1) establishing a 30-second regeneration step, 2) capturing CRH at 30 nM with a loading threshold of 0.12 nm, 3) loading 60 nM antibody (primary Ab) to saturate the immobilized peptide for 200 seconds, and 4) allowing the mixture of primary Ab and competing antibody from step 3 to bind for 200 seconds.

[0219] Octet data was processed with Fortebio's Data Analysis Software 11.0. After signal extraction, percent inhibition was calculated as follows: Inhibition rate (%) = (ab) / a * 100% For example, in the case of antibodies (Abn), Variant a: The signal of binding to the antigen by Abn alone (also called 100% signal of Abn) Variant b: If the Abn is a second Abn, it signals the binding of the second Abn to the antigen.

[0220] [Example 2] Generation of CRH knockout mice on the H2L2 Harbour Mice® background CRH knockout mice were prepared using the materials and methods described above on the H2L2 Harbour Mice® background. Figure 1 shows the coding sequence open reading frame of mouse CRH (GenBank NM_205769), along with the CRISPR targeting gRNA and PCR primers used. Specifically, the following primers were used for genotyping, i.e., for amplifying genomic DNA as depicted in Figure 1: CRHbp70F: 5'-GCC CTG CTC AGC AGG GGA TCC GT-3' (SEQ ID NO: 477) CRHbp215R: 5'-CTG TTG AGA TTC CCC AGG CGG AG-3' (SEQ ID NO: 478) CRHbp208F:5'-CTC AAC AGA AGT CCC GCT CGG-3' (SEQ ID NO: 479) CRHbp310R: 5'-GGA AAA AGT TAG CCG CAG CCT GG-3' (SEQ ID NO: 480)

[0221] The predicted PCR DNA fragment sizes for wild-type mice are 146 bp and 103 bp, respectively. PCR fragments smaller than these predicted sizes were shown to be deleted from the founder mice. Figures 2A-2B show the confirmation of CRH knockout founder mice using this method.

[0222] Figure 3 shows the location of an 11-bp deletion (5'-CAGCCGGTTCT-3') (SEQ ID NO: 465) in the CRH gene coding region 157-167, which leads to a downstream reading frameshift and a premature stop codon. This defective CRH open reading frame is unable to synthesize the CRH peptide. Founders with this deletion were selected for further breeding to homozygosity, rescue, and immunization.

[0223] [Example 3] Validation of anti-CRG mAb in CRH knockout mice on the H2L2 Harbour Mice® background Many positive hybridoma clones were identified from multiple hybridoma fusions by immunization of CRH peptides in CRH knockout mice on the H2L2 Harbour Mice® background. Monoclonal antibodies (mAbs) purified from hybridoma supernatants bound to CRH with high affinity in ELISA. However, three functional monoclonal antibody clones, hybridoma clones 16g2, 28b3, and 52g7, were confirmed to have blocking activity in a human CRHR1 luciferase reporter assay. These three clones, PR004289, PR004290, and PR006669, were recombinantly expressed with human IgG1 wild-type Fc, respectively. PR004289 (hybridoma clone 16g2), PR004290 (hybridoma clone 28b3), and PR006669 (hybridoma clone 52g7) share the same germline IGHV3-7 and IGKV2-28 sequences. PR004289 and PR004290 have one different residue at position 28 of the IGHV: I28 in PR004289 and T28 in PR004290, respectively. Both PR004289 and PR004290 have different complementarity-determining regions (CDRs) from PR006669. These three mAbs were designated Series 1 (PR004289 and PR004290) and Series 2 (PR006669) mAbs. See Table 2.

[0224] [Table 2]

[0225] [Example 4] Validation of anti-CRH mAb in wild-type mice More than 200,000 single B cells from wild-type mice immunized with CRH-KLH conjugates were screened using the Beacon system using the materials and methods described above. More than 500 monoclonal antibodies (mAbs) with binding activity in bead-binding assays were identified. More than 300 of these mAbs were recombinantly expressed with human IgG1 wild-type Fc. Many of these recombinantly expressed mAbs bound CRH with high affinity in ELISA. Eight series of functional mAbs with blocking activity in the human CRHR1 luciferase reporter assay were identified using the materials and methods described above. Different series of mAbs have different germline sequences and different CDRs. mAbs within each series share the same germline sequence but have different CDRs.

[0226] These mAbs were designated as series 3 to series 10 as follows: Series 3 includes PR301306. Series 4 includes PR301612. Series 5 includes PR301777, PR301788, PR301790, and PR301803. Series 6 includes PR301806. Series 7 includes PR302309, PR302332, and PR302333. Series 8 includes PR302315 and PR302335. Series 9 includes PR302328, PR302331, PR302334, and PR302339. Series 10 includes PR302312, PR302324, and PR302341.

[0227] See also Table 2 and Figures 4A-4N.

[0228] [Example 5] Antibody modification PTM removal of series 1 mAb PR004290 PR004290 contains two potential post-translational modifications (PTMs): DG in the heavy chain CDR2 (HCDR2) and NS in the light chain CDR1 (LCDR1). PR006669 has one potential PTM: DG in HCDR2. These potential PTMs were mutated individually or in various combinations in either HCDR2 alone, LCDR1 alone, or both, as detailed in Table 3. Their binding activity in CRH ELISA and functional blocking activity in human CRHR1 (hCRHR1), human CRHR2 (hCRHR2), mouse CRHR1 (mCRHR1), and mouse CRHR2 (mCRHR2) luciferase reporter assays are shown in Table 3 and Figures 5A–5L. Most ΔPTM mutants retained functional activity. PR005660 had a median inhibitory concentration (IC50) of approximately 10 nM in the hCRHR1 reporter assay, compared to approximately 5 nM for the parent PR004290.

[0229] [Table 3]

[0230] PTM removal of series 2 mAb PR006669 PR006745 and PR006746 are two ΔPTM mutants of PR006669, with somewhat weaker ELISA binding and function-blocking activities (see Table 4 and Figures 6A-6D).

[0231] [Table 4]

[0232] Humanization of Series 5 mAb PR301777 Fifteen humanized variants of PR301777 were recombinantly expressed and tested using the materials and methods described above. Many humanized variants maintained highly potent in vitro ELISA binding and function-blocking activity. See Table 5 and Figures 7A-7L.

[0233] [Table 5]

[0234] Humanization of Series 7 mAb PR302309 Twenty-one humanized variants of PR302309 were recombinantly expressed and tested using the materials and methods described above. Many humanized variants maintained highly potent in vitro ELISA binding and function-blocking activity. See Table 6 and Figures 8A-8D.

[0235] [Table 6]

[0236] Humanization of Series 9 mAb PR302334 Twenty-two humanized variants of PR302334 were recombinantly expressed and tested using the materials and methods described above. Many humanized variants maintained highly potent in vitro ELISA binding and function-blocking activity. See Table 7 and Figures 9A-9F.

[0237] [Table 7]

[0238] Humanization of Series 10 mAb PR302341 Twenty-seven humanized variants of PR302341 were recombinantly expressed and tested using the materials and methods described above. Many humanized variants maintained highly potent in vitro ELISA binding and function-blocking activity. See Table 8 and Figures 10A-10G.

[0239] [Table 8]

[0240] PTM removal of humanized series 10 mAbs PR302341-10, PR302341-20, and PR302341-23 Humanized series 10 mAbs PR302341-10, PR302341-20, and PR302341-23 were subjected to 4x4 double PTM removal mutagenesis using the materials and methods described above to generate a total of 48 mAbs (3x4x4), including the three parent humanized mAbs. Many humanized variants maintained highly potent in vitro ELISA binding and function-blocking activities. See Table 9 and Figures 11A-11L. PR302341-34, PR302341-48, PR302341-60, PR302341-28, and PR302341-55 were expanded for in vivo studies.

[0241] [Table 9]

[0242] [Example 6] Selectivity for human UCN1, human UCN2, and human UCN3 peptides All tested mAbs, except PR302334-26, bound to the human UCN1 peptide in ELISA assays. None of the tested mAbs bound to either the human UCN2 or human UCN3 peptides in ELISA assays. The binding affinity of these mAbs to human UCN1 was comparable to that of CRH by ELISA. However, only the PR302341 series mAb inhibited hUCN1-induced hCRHR1 luciferase reporter activity. In addition, these mAbs inhibited hUCN1-induced hCRHR1 reporter activity 30× to 267× less potently than CRH-induced hCRHR1 reporter activity. None of the mAbs blocked hUCN1-induced hCRHR2 luciferase reporter activity. See Table 10 and Figures 12A-12B, 13A-13D, and 14A-14H.

[0243] [Table 10]

[0244] [Example 7] Anti-CRH mAb binds to the N-terminal region of the CRH peptide To determine the binding region of the CRH peptide, the N-terminal region plus the central region, the central region alone, and the central region plus the C-terminal region of the CRH peptide were used for ELISA capture of CRH using the materials and methods described above. All mAbs bound to the N-terminal region plus the central region of the CRH peptide. Therefore, these mAbs likely bind to the N-terminal region of CRH. See Table 11 and Figures 15A-15D.

[0245] [Table 11]

[0246] [Example 8] Determining mAb affinity with Octet The binding affinity of the anti-CRH antibody was determined using the Octet platform using the materials and methods described above. The identified mAb has very high affinity, with equilibrium dissociation constant (KD) values ​​of approximately single to double digit pM. See Table 12.

[0247] [Table 12]

[0248] [Example 9] Binding of antibody-binding epitopes by ELISA The selected mAbs were tested for their ability to block each other's binding to biotin-CRH in a paired competitive ELISA using the materials and methods described above. Table 13 shows that these mAbs blocked each other's binding to CRH, thus indicating that they likely bind to similar epitopes on the N-terminus of CRH.

[0249] [Table 13]

[0250] [Example 10] Octet binding of antibody-binding epitopes The selected mAbs were tested for their ability to block each other's binding to biotin-CRH in a paired competitive Octet assay using the materials and methods described above. Figure 16 shows a diagram of the competitive assay using the Octet platform. Table 14 shows that these mAbs blocked each other's binding to CRH, thus indicating the possibility that they bind to similar epitopes, possibly at the N-terminus of CRH.

[0251] [Table 14]

[0252] [Example 11] In vivo efficacy studies Restraint-induced transient increases in ACTH and corticosterone in wild-type mice Treatment with anti-CRH mAbs PR004290 (series 1 mAb), PR006152 (series 1 mAb), and PR302050 (series 5 mAb) (20 mg / kg IP) significantly inhibited adrenocorticotropic hormone (ACTH) and corticosterone concentrations in wild-type mice exposed to the restraint-induced stress model discussed above. One day after mAb administration, plasma ACTH and corticosterone concentrations were inhibited to basal, non-stressed levels. Fourteen days after a single dose of mAb, ACTH and corticosterone concentrations remained partially inhibited. See Figure 17.

[0253] A model of constitutively high ACTH in Mrap1KO mice Mrap1KO mice had constitutively elevated plasma ACTH concentrations of approximately 5,000 pg / ml, approximately 50-fold higher than the basal ACTH concentration in wild-type mice and approximately 10-fold higher than that observed with restraint-induced stress in wild-type mice. A single 20 mg / kg dose of the anti-CRH mAb PR302050 (series 5 mAb) significantly reduced the constitutively elevated ACTH concentrations in Mrap1KO mice both 2 and 14 days after administration. See Figure 18.

[0254] A single dose of the anti-CRH mAb PR302064 (series 5 mAb) significantly reduced the constitutively high ACTH levels in Mrap1KO mice at 20 mg / kg, 5 mg / kg, and 1 mg / kg doses on days 2, 16, and 28, respectively. PR302064 at 20 mg / kg reduced ACTH by 73%, 82%, and 55% on days 2, 16, and 28, respectively. PR302064 at 5 mg / kg reduced ACTH by 74%, 45%, and 34% on days 2, 16, and 28, respectively. PR302064 at 1 mg / kg reduced ACTH by 47%, 31%, and 23% on days 2, 16, and 28, respectively. Other mAbs, including PR302050 (series 5 mAb), PR005660 (series 1 mAb), and PR006669 (series 2 mAb), also significantly reduced ACTH on days 2 and 16 after dosing at 5 mg / kg. See Figures 19A-19C.

[0255] Additional anti-CRH mAbs were tested in the Mrap1KO model. A single 20 mg / kg dose of PR302334-26 (series 9 mAb), PR302341-28, PR302341-34, PR302341-55, and PR302341-60 (all series 10 mAbs) also significantly reduced plasma ACTH concentrations in Mrap1KO mice. See Figures 20A-20B.

[0256] PR302038, PR005660, and PR302050 dose-dependently reduced plasma ACTH concentrations in Mrap1KO mice at antibody doses of 20 mg / kg and 5 mg / kg. See Figures 21A-21B.

[0257] PR302334-24 and PR005660 significantly reduced plasma ACTH concentrations in Mrap1KO mice at 20 mg / kg, whereas the anti-CRH mAb PR301429, which has comparable binding affinity but is a non-blocker in vitro, did not inhibit ACTH production. PR303394, an analog of CTRND05, was used as a control. See Figures 22A-22B.

[0258] Anti-CRH mAb PR302064 had superior efficacy to SSR125543A in the restraint-induced ACTH model in wild-type mice The anti-CRH mAb PR302064 (series 5 mAb) at 20 mg / kg was more effective than the CRHR1 small molecule antagonist SSR125543A (Clinecelfont) at 30 mg / kg in reducing restraint-induced ACTH in wild-type mice. PR302064 had a more complete inhibition of both ACTH and corticosterone and a longer duration of action. The effect of this mAb lasted for at least 16 days. See Figures 23A-23B.

[0259] Anti-CRH mAbs PR302064 and PR302334-24 were more effective than SSR125543A in a model of constitutively elevated ACTH in Marp1KO mice The anti-CRH mAb PR302064 at 20 mg / kg and PR302334-24 at 20 mg / kg, 10 mg / kg, and 5 mg / kg were all more effective than the CRHR1 small molecule antagonist SSR125543A (Clinecelfont) at 30 mg / kg in reducing plasma ACTH levels in a model of constitutively elevated ACTH in Marp1KO mice. PR302064 and PR302334-24 had more complete inhibition and, more importantly, a longer duration of action. The effects of these two mAbs lasted for at least 14 days. See Figures 24A-24B.

[0260] Anti-CRH mAb PR302334-24 was more effective than SSR125543A in the restraint stress-induced high ACTH and corticosterone model in wild-type mice Both 20 mg / kg and 5 mg / kg doses of the anti-CRH mAb PR302334-24 were more effective than 30 mg / kg of the CRHR1 small molecule antagonist SSR125543A (Clinecelfont) in reducing plasma ACTH and corticosterone levels in the wild-type mouse restraint stress-induced anti-ACTH and corticosterone model. PR302334-24 had more complete inhibition and, more importantly, a longer duration of action. The effect of PR302334-24 lasted for at least 17 days. See Figures 25A-25B.

[0261] In vivo PK studies mAbs PR004290, PR302050, and PR302064 were tested in C57BL / 6 wild-type mice at a single IV dose of 5 mg / kg. Specifically, anti-CRH mAbs in human IgG1WT-Fc present in mouse serum were captured with a goat anti-human Fc polyclonal antibody and detected with a goat anti-human (H+L) secondary antibody conjugated to HRP. Figure 26A shows a diagram of this method.

[0262] As shown in Table 15 and Figures 26B to 26D, the serum half-lives (T 1 / 2 ) were 14, 21, and 18 days, respectively. These data indicate that these mAbs have a typically human IgG1 PK profile, making them desirable for clinical application. 1 / 2 It indicates that it has.

[0263] [Table 15]

[0264] All publications, patents, and patent applications mentioned in this application are incorporated herein by reference in their entirety to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated by reference herein. In addition, citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent section headings are used, they should not be construed as necessarily limiting.

Claims

1. An antibody or antigen-binding fragment thereof that specifically binds to corticotropin-releasing hormone (CRH), (a) a heavy chain variable domain (VH) CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 22-35, a VH CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 53-78, and a VH CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 113-122, or the amino acid sequence of a PDV or GID; and (b) a light chain variable domain (VL) CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 152-174, a VL CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 193-198, and a VL CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 223-232. An antibody or antigen-binding fragment thereof comprising:

2. the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:53, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:152, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:54, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:152, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:55, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:152, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:53, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:153, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:54, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:153, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:55, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:153, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:54, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:154, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:54, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:155, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:54, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:156, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:54, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:157, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:55, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:154, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:55, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:155, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:55, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:156, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:55, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:157, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:56, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:154, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:56, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:155, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:56, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:156, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:56, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:157, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:223; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:23, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:57, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:114, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:158, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:224; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:23, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:58, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:114, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:158, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:224; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:23, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:59, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:114, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:158, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:224; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:23, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:60, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:114, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:158, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:224; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:24, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:61, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:115, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:159, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:194, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:225; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:25, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:62, the VH CDR3 comprises the amino acid sequence of PDV, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:160, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:195, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:226; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:24, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:63, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:115, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:161, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:196, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:225; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:26, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:64, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:116, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:162, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:227; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:24, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:63, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:115, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:163, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:196, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:225; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:24, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:65, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:115, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:161, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:196, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:225; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:27, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:66, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:117, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:164, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:228; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:28, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:67, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:118, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:165, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:227; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:68, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:119, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:166, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:227; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:30, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:69, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:120, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:167, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:229; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:70, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:166, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 31, the VH CDR2 comprises the amino acid sequence of SEQ ID NO: 71, the VH CDR3 comprises the amino acid sequence of GID, the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 168, the VL CDR2 comprises the amino acid sequence of SEQ ID NO: 198, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO: 231; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 32, the VH CDR2 comprises the amino acid sequence of SEQ ID NO: 72, the VH CDR3 comprises the amino acid sequence of SEQ ID NO: 118, the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 166, the VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO: 227; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 33, the VH CDR2 comprises the amino acid sequence of SEQ ID NO: 73, the VH CDR3 comprises the amino acid sequence of SEQ ID NO: 118, the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 169, the VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO: 227; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 34, the VH CDR2 comprises the amino acid sequence of SEQ ID NO: 71, the VH CDR3 comprises the amino acid sequence of GID, the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 168, the VL CDR2 comprises the amino acid sequence of SEQ ID NO: 198, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO: 231; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 30, the VH CDR2 comprises the amino acid sequence of SEQ ID NO: 74, the VH CDR3 comprises the amino acid sequence of SEQ ID NO: 122, the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 170, the VL CDR2 comprises the amino acid sequence of SEQ ID NO: 197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO: 232; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 35, the VH CDR2 comprises the amino acid sequence of SEQ ID NO: 71, the VH CDR3 comprises the amino acid sequence of GID, the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 168, the VL CDR2 comprises the amino acid sequence of SEQ ID NO: 198, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO: 231; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:75, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:171, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:76, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:171, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:77, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:171, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:75, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:172, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:75, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:173, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:76, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:172, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:77, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:173, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:76, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:173, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:77, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:172, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:78, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:172, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:78, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:173, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:76, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:174, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:77, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:174, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:78, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:174, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:78, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:171, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO:29, the VH CDR2 comprises the amino acid sequence of SEQ ID NO:75, the VH CDR3 comprises the amino acid sequence of SEQ ID NO:121, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:174, the VL CDR2 comprises the amino acid sequence of SEQ ID NO:197, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO:230; or the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 22, the VH CDR2 comprises the amino acid sequence of SEQ ID NO: 53, the VH CDR3 comprises the amino acid sequence of SEQ ID NO: 113, the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 152, the VL CDR2 comprises the amino acid sequence of SEQ ID NO: 193, and the VL CDR3 comprises the amino acid sequence of SEQ ID NO: 223; The antibody or antigen-binding fragment thereof of claim 1. (a) a VH comprising an amino acid sequence having at least 90% identity to any one of SEQ ID NOs: 240 to 295, 459, and 461; and / or (b) a VL comprising an amino acid sequence having at least 90% identity to any one of SEQ ID NOs: 296-347 and 462; The antibody or antigen-binding fragment thereof of claim 1, comprising: (a) a VH comprising any one of the amino acid sequences of SEQ ID NOs: 240 to 295, 459, and 461; and / or (b) a VL comprising any one of the amino acid sequences of SEQ ID NOs: 296-347 and 462; The antibody or antigen-binding fragment thereof of claim 3, comprising: Claim 5: The VH comprises the amino acid sequence of SEQ ID NO: 240, and / or the VL comprises the amino acid sequence of SEQ ID NO: 296, or the VH comprises the amino acid sequence of SEQ ID NO: 241, and / or the VL comprises the amino acid sequence of SEQ ID NO: 296, or the VH comprises the amino acid sequence of SEQ ID NO: 242, and / or the VL comprises the amino acid sequence of SEQ ID NO: 296, or the VH comprises the amino acid sequence of SEQ ID NO: 240, and / or the VL comprises the amino acid sequence of SEQ ID NO: 297, or the VH comprises the amino acid sequence of SEQ ID NO: 241, and / or the VL comprises the amino acid sequence of SEQ ID NO: 297, or the VH comprises the amino acid sequence of SEQ ID NO: 242, and / or the VL comprises the amino acid sequence of SEQ ID NO: 297, or the VH comprises the amino acid sequence of SEQ ID NO: 241, and / or the VL comprises the amino acid sequence of SEQ ID NO: 298, or the VH comprises the amino acid sequence of SEQ ID NO: 241 and the VL comprises the amino acid sequence of SEQ ID NO: 299; or the VH comprises the amino acid sequence of SEQ ID NO: 241 and the VL comprises the amino acid sequence of SEQ ID NO: 300; or the VH comprises the amino acid sequence of SEQ ID NO: 241, and / or the VL comprises the amino acid sequence of SEQ ID NO: 301, or the VH comprises the amino acid sequence of SEQ ID NO: 242, and / or the VL comprises the amino acid sequence of SEQ ID NO: 298, or the VH comprises the amino acid sequence of SEQ ID NO: 242, and / or the VL comprises the amino acid sequence of SEQ ID NO: 299, or the VH comprises the amino acid sequence of SEQ ID NO: 242, and / or the VL comprises the amino acid sequence of SEQ ID NO: 300, or the VH comprises the amino acid sequence of SEQ ID NO: 242, and / or the VL comprises the amino acid sequence of SEQ ID NO: 301, or the VH comprises the amino acid sequence of SEQ ID NO: 243, and / or the VL comprises the amino acid sequence of SEQ ID NO: 298, or the VH comprises the amino acid sequence of SEQ ID NO: 243, and / or the VL comprises the amino acid sequence of SEQ ID NO: 299, or the VH comprises the amino acid sequence of SEQ ID NO: 243, and / or the VL comprises the amino acid sequence of SEQ ID NO: 300, or the VH comprises the amino acid sequence of SEQ ID NO: 243, and / or the VL comprises the amino acid sequence of SEQ ID NO: 301, or the VH comprises the amino acid sequence of SEQ ID NO: 244, and / or the VL comprises the amino acid sequence of SEQ ID NO: 302, or the VH comprises the amino acid sequence of SEQ ID NO: 245, and / or the VL comprises the amino acid sequence of SEQ ID NO: 302, or the VH comprises the amino acid sequence of SEQ ID NO: 246, and / or the VL comprises the amino acid sequence of SEQ ID NO: 302, or the VH comprises the amino acid sequence of SEQ ID NO: 247, and / or the VL comprises the amino acid sequence of SEQ ID NO: 302, or the VH comprises the amino acid sequence of SEQ ID NO: 248, and / or the VL comprises the amino acid sequence of SEQ ID NO: 303, or the VH comprises the amino acid sequence of SEQ ID NO: 249, and / or the VL comprises the amino acid sequence of SEQ ID NO: 304, or the VH comprises the amino acid sequence of SEQ ID NO: 250, and / or the VL comprises the amino acid sequence of SEQ ID NO: 305, or the VH comprises the amino acid sequence of SEQ ID NO: 251, and / or the VL comprises the amino acid sequence of SEQ ID NO: 306, or the VH comprises the amino acid sequence of SEQ ID NO: 252, and / or the VL comprises the amino acid sequence of SEQ ID NO: 305, or the VH comprises the amino acid sequence of SEQ ID NO: 252, and / or the VL comprises the amino acid sequence of SEQ ID NO: 307, or the VH comprises the amino acid sequence of SEQ ID NO: 253, and / or the VL comprises the amino acid sequence of SEQ ID NO: 308, or the VH comprises the amino acid sequence of SEQ ID NO: 254, and / or the VL comprises the amino acid sequence of SEQ ID NO: 309, or the VH comprises the amino acid sequence of SEQ ID NO: 255, and / or the VL comprises the amino acid sequence of SEQ ID NO: 310, or the VH comprises the amino acid sequence of SEQ ID NO: 255, and / or the VL comprises the amino acid sequence of SEQ ID NO: 311, or the VH comprises the amino acid sequence of SEQ ID NO: 255, and / or the VL comprises the amino acid sequence of SEQ ID NO: 312, or the VH comprises the amino acid sequence of SEQ ID NO: 256, and / or the VL comprises the amino acid sequence of SEQ ID NO: 310, or the VH comprises the amino acid sequence of SEQ ID NO: 256, and / or the VL comprises the amino acid sequence of SEQ ID NO: 311, or the VH comprises the amino acid sequence of SEQ ID NO: 256, and / or the VL comprises the amino acid sequence of SEQ ID NO: 312, or the VH comprises the amino acid sequence of SEQ ID NO: 257, and / or the VL comprises the amino acid sequence of SEQ ID NO: 310, or the VH comprises the amino acid sequence of SEQ ID NO: 257, and / or the VL comprises the amino acid sequence of SEQ ID NO: 311, or the VH comprises the amino acid sequence of SEQ ID NO: 257, and / or the VL comprises the amino acid sequence of SEQ ID NO: 312, or the VH comprises the amino acid sequence of SEQ ID NO: 258, and / or the VL comprises the amino acid sequence of SEQ ID NO: 310, or the VH comprises the amino acid sequence of SEQ ID NO: 258, and / or the VL comprises the amino acid sequence of SEQ ID NO: 311, or the VH comprises the amino acid sequence of SEQ ID NO: 258, and / or the VL comprises the amino acid sequence of SEQ ID NO: 312, or the VH comprises the amino acid sequence of SEQ ID NO: 259, and / or the VL comprises the amino acid sequence of SEQ ID NO: 310, or the VH comprises the amino acid sequence of SEQ ID NO: 259, and / or the VL comprises the amino acid sequence of SEQ ID NO: 311, or the VH comprises the amino acid sequence of SEQ ID NO: 259, and / or the VL comprises the amino acid sequence of SEQ ID NO: 312, or the VH comprises the amino acid sequence of SEQ ID NO: 260, and / or the VL comprises the amino acid sequence of SEQ ID NO: 317, or the VH comprises the amino acid sequence of SEQ ID NO: 260, and / or the VL comprises the amino acid sequence of SEQ ID NO: 313, or the VH comprises the amino acid sequence of SEQ ID NO: 261, and / or the VL comprises the amino acid sequence of SEQ ID NO: 314, or the VH comprises the amino acid sequence of SEQ ID NO: 261, and / or the VL comprises the amino acid sequence of SEQ ID NO: 315, or the VH comprises the amino acid sequence of SEQ ID NO: 261, and / or the VL comprises the amino acid sequence of SEQ ID NO: 316, or the VH comprises the amino acid sequence of SEQ ID NO: 262, and / or the VL comprises the amino acid sequence of SEQ ID NO: 317, or the VH comprises the amino acid sequence of SEQ ID NO: 261, and / or the VL comprises the amino acid sequence of SEQ ID NO: 317, or the VH comprises the amino acid sequence of SEQ ID NO: 263, and / or the VL comprises the amino acid sequence of SEQ ID NO: 317, or the VH comprises the amino acid sequence of SEQ ID NO: 264, and / or the VL comprises the amino acid sequence of SEQ ID NO: 317, or the VH comprises the amino acid sequence of SEQ ID NO: 262, and / or the VL comprises the amino acid sequence of SEQ ID NO: 318, or the VH comprises the amino acid sequence of SEQ ID NO: 263, and / or the VL comprises the amino acid sequence of SEQ ID NO: 318, or the VH comprises the amino acid sequence of SEQ ID NO: 264, and / or the VL comprises the amino acid sequence of SEQ ID NO: 318, or the VH comprises the amino acid sequence of SEQ ID NO: 260, and / or the VL comprises the amino acid sequence of SEQ ID NO: 318, or the VH comprises the amino acid sequence of SEQ ID NO: 264, and / or the VL comprises the amino acid sequence of SEQ ID NO: 314, or the VH comprises the amino acid sequence of SEQ ID NO: 264, and / or the VL comprises the amino acid sequence of SEQ ID NO: 316, or the VH comprises the amino acid sequence of SEQ ID NO: 260, and / or the VL comprises the amino acid sequence of SEQ ID NO: 319, or the VH comprises the amino acid sequence of SEQ ID NO: 260, and / or the VL comprises the amino acid sequence of SEQ ID NO: 314, or the VH comprises the amino acid sequence of SEQ ID NO: 260, and / or the VL comprises the amino acid sequence of SEQ ID NO: 315, or the VH comprises the amino acid sequence of SEQ ID NO: 260, and / or the VL comprises the amino acid sequence of SEQ ID NO: 316, or the VH comprises the amino acid sequence of SEQ ID NO: 261, and / or the VL comprises the amino acid sequence of SEQ ID NO: 313, or the VH comprises the amino acid sequence of SEQ ID NO: 261, and / or the VL comprises the amino acid sequence of SEQ ID NO: 318, or the VH comprises the amino acid sequence of SEQ ID NO: 261, and / or the VL comprises the amino acid sequence of SEQ ID NO: 319, or the VH comprises the amino acid sequence of SEQ ID NO: 265, and / or the VL comprises the amino acid sequence of SEQ ID NO: 320, or the VH comprises the amino acid sequence of SEQ ID NO: 266, and / or the VL comprises the amino acid sequence of SEQ ID NO: 321, or the VH comprises the amino acid sequence of SEQ ID NO: 267, and / or the VL comprises the amino acid sequence of SEQ ID NO: 322, or the VH comprises the amino acid sequence of SEQ ID NO: 268, and / or the VL comprises the amino acid sequence of SEQ ID NO: 323, or the VH comprises the amino acid sequence of SEQ ID NO: 269, and / or the VL comprises the amino acid sequence of SEQ ID NO: 324, or the VH comprises the amino acid sequence of SEQ ID NO: 270, and / or the VL comprises the amino acid sequence of SEQ ID NO: 325, or the VH comprises the amino acid sequence of SEQ ID NO: 271, and / or the VL comprises the amino acid sequence of SEQ ID NO: 326, or the VH comprises the amino acid sequence of SEQ ID NO: 272, and / or the VL comprises the amino acid sequence of SEQ ID NO: 331, or the VH comprises the amino acid sequence of SEQ ID NO: 272, and / or the VL comprises the amino acid sequence of SEQ ID NO: 327, or the VH comprises the amino acid sequence of SEQ ID NO: 273, and / or the VL comprises the amino acid sequence of SEQ ID NO: 328, or the VH comprises the amino acid sequence of SEQ ID NO: 273, and / or the VL comprises the amino acid sequence of SEQ ID NO: 329, or the VH comprises the amino acid sequence of SEQ ID NO: 273, and / or the VL comprises the amino acid sequence of SEQ ID NO: 330, or the VH comprises the amino acid sequence of SEQ ID NO: 273, and / or the VL comprises the amino acid sequence of SEQ ID NO: 331, or the VH comprises the amino acid sequence of SEQ ID NO: 274, and / or the VL comprises the amino acid sequence of SEQ ID NO: 331, or the VH comprises the amino acid sequence of SEQ ID NO: 275, and / or the VL comprises the amino acid sequence of SEQ ID NO: 331, or the VH comprises the amino acid sequence of SEQ ID NO: 276, and / or the VL comprises the amino acid sequence of SEQ ID NO: 331, or the VH comprises the amino acid sequence of SEQ ID NO: 277, and / or the VL comprises the amino acid sequence of SEQ ID NO: 331, or the VH comprises the amino acid sequence of SEQ ID NO: 272, and / or the VL comprises the amino acid sequence of SEQ ID NO: 332, or the VH comprises the amino acid sequence of SEQ ID NO: 276, and / or the VL comprises the amino acid sequence of SEQ ID NO: 332, or the VH comprises the amino acid sequence of SEQ ID NO: 275, and / or the VL comprises the amino acid sequence of SEQ ID NO: 328, or the VH comprises the amino acid sequence of SEQ ID NO: 275, and / or the VL comprises the amino acid sequence of SEQ ID NO: 330, or the VH comprises the amino acid sequence of SEQ ID NO: 277, and / or the VL comprises the amino acid sequence of SEQ ID NO: 328, or the VH comprises the amino acid sequence of SEQ ID NO: 277, and / or the VL comprises the amino acid sequence of SEQ ID NO: 330, or the VH comprises the amino acid sequence of SEQ ID NO: 272, and / or the VL comprises the amino acid sequence of SEQ ID NO: 333, or the VH comprises the amino acid sequence of SEQ ID NO: 272, and / or the VL comprises the amino acid sequence of SEQ ID NO: 328, or the VH comprises the amino acid sequence of SEQ ID NO: 272, and / or the VL comprises the amino acid sequence of SEQ ID NO: 329, or the VH comprises the amino acid sequence of SEQ ID NO: 272, and / or the VL comprises the amino acid sequence of SEQ ID NO: 330, or the VH comprises the amino acid sequence of SEQ ID NO: 273, and / or the VL comprises the amino acid sequence of SEQ ID NO: 327, or the VH comprises the amino acid sequence of SEQ ID NO: 273, and / or the VL comprises the amino acid sequence of SEQ ID NO: 332, or the VH comprises the amino acid sequence of SEQ ID NO: 273, and / or the VL comprises the amino acid sequence of SEQ ID NO: 333, or the VH comprises the amino acid sequence of SEQ ID NO: 278, and / or the VL comprises the amino acid sequence of SEQ ID NO: 334, or the VH comprises the amino acid sequence of SEQ ID NO: 279, and / or the VL comprises the amino acid sequence of SEQ ID NO: 323, or the VH comprises the amino acid sequence of SEQ ID NO: 280, and / or the VL comprises the amino acid sequence of SEQ ID NO: 339, or the VH comprises the amino acid sequence of SEQ ID NO: 280, and / or the VL comprises the amino acid sequence of SEQ ID NO: 335, or the VH comprises the amino acid sequence of SEQ ID NO: 281, and / or the VL comprises the amino acid sequence of SEQ ID NO: 336, or the VH comprises the amino acid sequence of SEQ ID NO: 281, and / or the VL comprises the amino acid sequence of SEQ ID NO: 337, or the VH comprises the amino acid sequence of SEQ ID NO: 281, and / or the VL comprises the amino acid sequence of SEQ ID NO: 338, or the VH comprises the amino acid sequence of SEQ ID NO: 282, and / or the VL comprises the amino acid sequence of SEQ ID NO: 339, or the VH comprises the amino acid sequence of SEQ ID NO: 281, and / or the VL comprises the amino acid sequence of SEQ ID NO: 339, or the VH comprises the amino acid sequence of SEQ ID NO: 283, and / or the VL comprises the amino acid sequence of SEQ ID NO: 339, or the VH comprises the amino acid sequence of SEQ ID NO: 284, and / or the VL comprises the amino acid sequence of SEQ ID NO: 339, or the VH comprises the amino acid sequence of SEQ ID NO: 285, and / or the VL comprises the amino acid sequence of SEQ ID NO: 339, or the VH comprises the amino acid sequence of SEQ ID NO: 286, and / or the VL comprises the amino acid sequence of SEQ ID NO: 339, or the VH comprises the amino acid sequence of SEQ ID NO: 282, and / or the VL comprises the amino acid sequence of SEQ ID NO: 340, or the VH comprises the amino acid sequence of SEQ ID NO: 280, and / or the VL comprises the amino acid sequence of SEQ ID NO: 340, or the VH comprises the amino acid sequence of SEQ ID NO: 283, and / or the VL comprises the amino acid sequence of SEQ ID NO: 340, or the VH comprises the amino acid sequence of SEQ ID NO: 284, and / or the VL comprises the amino acid sequence of SEQ ID NO: 340, or the VH comprises the amino acid sequence of SEQ ID NO: 285, and / or the VL comprises the amino acid sequence of SEQ ID NO: 340, or the VH comprises the amino acid sequence of SEQ ID NO: 286, and / or the VL comprises the amino acid sequence of SEQ ID NO: 340, or the VH comprises the amino acid sequence of SEQ ID NO: 284, and / or the VL comprises the amino acid sequence of SEQ ID NO: 336, or the VH comprises the amino acid sequence of SEQ ID NO: 284, and / or the VL comprises the amino acid sequence of SEQ ID NO: 338, or the VH comprises the amino acid sequence of SEQ ID NO: 286, and / or the VL comprises the amino acid sequence of SEQ ID NO: 336, or the VH comprises the amino acid sequence of SEQ ID NO: 286, and / or the VL comprises the amino acid sequence of SEQ ID NO: 338, or the VH comprises the amino acid sequence of SEQ ID NO: 287, and / or the VL comprises the amino acid sequence of SEQ ID NO: 340, or the VH comprises the amino acid sequence of SEQ ID NO: 288, and / or the VL comprises the amino acid sequence of SEQ ID NO: 340, or the VH comprises the amino acid sequence of SEQ ID NO: 280, and / or the VL comprises the amino acid sequence of SEQ ID NO: 341, or the VH comprises the amino acid sequence of SEQ ID NO: 286, and / or the VL comprises the amino acid sequence of SEQ ID NO: 342, or the VH comprises the amino acid sequence of SEQ ID NO: 286, and / or the VL comprises the amino acid sequence of SEQ ID NO: 343, or the VH comprises the amino acid sequence of SEQ ID NO: 287, and / or the VL comprises the amino acid sequence of SEQ ID NO: 342, or the VH comprises the amino acid sequence of SEQ ID NO: 288, and / or the VL comprises the amino acid sequence of SEQ ID NO: 343, or the VH comprises the amino acid sequence of SEQ ID NO: 287, and / or the VL comprises the amino acid sequence of SEQ ID NO: 343, or the VH comprises the amino acid sequence of SEQ ID NO: 288, and / or the VL comprises the amino acid sequence of SEQ ID NO: 342, or the VH comprises the amino acid sequence of SEQ ID NO: 289, and / or the VL comprises the amino acid sequence of SEQ ID NO: 342, or the VH comprises the amino acid sequence of SEQ ID NO: 289, and / or the VL comprises the amino acid sequence of SEQ ID NO: 343, or the VH comprises the amino acid sequence of SEQ ID NO: 287, and / or the VL comprises the amino acid sequence of SEQ ID NO: 344, or the VH comprises the amino acid sequence of SEQ ID NO: 288, and / or the VL comprises the amino acid sequence of SEQ ID NO: 344, or the VH comprises the amino acid sequence of SEQ ID NO: 280, and / or the VL comprises the amino acid sequence of SEQ ID NO: 336, or the VH comprises the amino acid sequence of SEQ ID NO: 289, and / or the VL comprises the amino acid sequence of SEQ ID NO: 344, or the VH comprises the amino acid sequence of SEQ ID NO: 289, and / or the VL comprises the amino acid sequence of SEQ ID NO: 340, or the VH comprises the amino acid sequence of SEQ ID NO: 286, and / or the VL comprises the amino acid sequence of SEQ ID NO: 344, or the VH comprises the amino acid sequence of SEQ ID NO: 290, and / or the VL comprises the amino acid sequence of SEQ ID NO: 336, or the VH comprises the amino acid sequence of SEQ ID NO: 291, and / or the VL comprises the amino acid sequence of SEQ ID NO: 336, or the VH comprises the amino acid sequence of SEQ ID NO: 292, and / or the VL comprises the amino acid sequence of SEQ ID NO: 336, or the VH comprises the amino acid sequence of SEQ ID NO: 290, and / or the VL comprises the amino acid sequence of SEQ ID NO: 345, or the VH comprises the amino acid sequence of SEQ ID NO: 291, and / or the VL comprises the amino acid sequence of SEQ ID NO: 345, or the VH comprises the amino acid sequence of SEQ ID NO: 292, and / or the VL comprises the amino acid sequence of SEQ ID NO: 345, or the VH comprises the amino acid sequence of SEQ ID NO: 290, and / or the VL comprises the amino acid sequence of SEQ ID NO: 346, or the VH comprises the amino acid sequence of SEQ ID NO: 280, and / or the VL comprises the amino acid sequence of SEQ ID NO: 337, or the VH comprises the amino acid sequence of SEQ ID NO: 291, and / or the VL comprises the amino acid sequence of SEQ ID NO: 346, or the VH comprises the amino acid sequence of SEQ ID NO: 292, and / or the VL comprises the amino acid sequence of SEQ ID NO: 346, or the VH comprises the amino acid sequence of SEQ ID NO: 290, and / or the VL comprises the amino acid sequence of SEQ ID NO: 347, or the VH comprises the amino acid sequence of SEQ ID NO: 291, and / or the VL comprises the amino acid sequence of SEQ ID NO: 347, or the VH comprises the amino acid sequence of SEQ ID NO: 292, and / or the VL comprises the amino acid sequence of SEQ ID NO: 347, or the VH comprises the amino acid sequence of SEQ ID NO: 293, and / or the VL comprises the amino acid sequence of SEQ ID NO: 340, or the VH comprises the amino acid sequence of SEQ ID NO: 294, and / or the VL comprises the amino acid sequence of SEQ ID NO: 340, or the VH comprises the amino acid sequence of SEQ ID NO: 295, and / or the VL comprises the amino acid sequence of SEQ ID NO: 340, or the VH comprises the amino acid sequence of SEQ ID NO: 293, and / or the VL comprises the amino acid sequence of SEQ ID NO: 343, or the VH comprises the amino acid sequence of SEQ ID NO: 294, and / or the VL comprises the amino acid sequence of SEQ ID NO: 343, or the VH comprises the amino acid sequence of SEQ ID NO: 280, and / or the VL comprises the amino acid sequence of SEQ ID NO: 338, or the VH comprises the amino acid sequence of SEQ ID NO: 295, and / or the VL comprises the amino acid sequence of SEQ ID NO: 343, or the VH comprises the amino acid sequence of SEQ ID NO: 293, and / or the VL comprises the amino acid sequence of SEQ ID NO: 342, or the VH comprises the amino acid sequence of SEQ ID NO: 294, and / or the VL comprises the amino acid sequence of SEQ ID NO: 342, or the VH comprises the amino acid sequence of SEQ ID NO: 295, and / or the VL comprises the amino acid sequence of SEQ ID NO: 342, or the VH comprises the amino acid sequence of SEQ ID NO: 293, and / or the VL comprises the amino acid sequence of SEQ ID NO: 344, or the VH comprises the amino acid sequence of SEQ ID NO: 294, and / or the VL comprises the amino acid sequence of SEQ ID NO: 344, or the VH comprises the amino acid sequence of SEQ ID NO: 295, and / or the VL comprises the amino acid sequence of SEQ ID NO: 344, or the VH comprises the amino acid sequence of SEQ ID NO: 281, and / or the VL comprises the amino acid sequence of SEQ ID NO: 345, or the VH comprises the amino acid sequence of SEQ ID NO: 281, and / or the VL comprises the amino acid sequence of SEQ ID NO: 346, or the VH comprises the amino acid sequence of SEQ ID NO: 281, and / or the VL comprises the amino acid sequence of SEQ ID NO: 347, or the VH comprises the amino acid sequence of SEQ ID NO: 281, and / or the VL comprises the amino acid sequence of SEQ ID NO: 335, or the VH comprises the amino acid sequence of SEQ ID NO: 283, and / or the VL comprises the amino acid sequence of SEQ ID NO: 343, or the VH comprises the amino acid sequence of SEQ ID NO: 283, and / or the VL comprises the amino acid sequence of SEQ ID NO: 342, or the VH comprises the amino acid sequence of SEQ ID NO: 283, and / or the VL comprises the amino acid sequence of SEQ ID NO: 344, or the VH comprises the amino acid sequence of SEQ ID NO: 281, and / or the VL comprises the amino acid sequence of SEQ ID NO: 340, or the VH comprises the amino acid sequence of SEQ ID NO: 281, and / or the VL comprises the amino acid sequence of SEQ ID NO: 341, or the VH comprises the amino acid sequence of SEQ ID NO: 461, and / or the VL comprises the amino acid sequence of SEQ ID NO: 462, or wherein the VH comprises the amino acid sequence of SEQ ID NO: 459, and / or the VL comprises the amino acid sequence of SEQ ID NO: 296; The antibody or antigen-binding fragment thereof according to claim 4. (a) a heavy chain comprising an amino acid sequence having at least 90% identity to any one of SEQ ID NOs: 348-403, 460, and 463; and / or (b) a light chain comprising an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 404-455 and 464. The antibody or antigen-binding fragment thereof of claim 1, comprising: (a) a heavy chain comprising any one of the amino acid sequences set forth in SEQ ID NOs: 348 to 403, 460, and 463; and / or (b) a light chain comprising the amino acid sequence of any one of SEQ ID NOs: 404-455 and 464. The antibody or antigen-binding fragment thereof of claim 6, comprising:

8. The heavy chain comprises the amino acid sequence of SEQ ID NO: 348, and / or the light chain comprises the amino acid sequence of SEQ ID NO: 404, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 349 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 404, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 350 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 404, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 348 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 405, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 349 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 405, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 350 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 405, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 349 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 406, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 349 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 407, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 349 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 408, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 349 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 409, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 350 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 406, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 350 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 407, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 350 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 408, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 350 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 409, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 351 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 406, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 351 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 407, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 351 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 408, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 351 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 409, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 352 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 410, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 353 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 410, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 354 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 410, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 355 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 410, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 356 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 411, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 357 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 412, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 358 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 413, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 359 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 414, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 360, and / or the light chain comprises the amino acid sequence of SEQ ID NO: 413, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 360 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 415, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 361 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 416, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 362 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 417, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 363 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 418, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 363 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 419, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 363 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 420, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 364 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 418, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 364 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 419, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 364 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 420, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 365 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 418, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 365 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 419, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 365 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 420, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 366 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 418, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 366 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 419, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 366 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 420, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 367 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 418, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 367 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 419, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 367 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 420, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 368 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 425, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 368 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 421, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 369 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 422, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 369 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 423, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 369 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 424, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 370 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 425, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 369 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 425, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 371 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 425, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 372 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 425, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 370 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 426, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 371 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 426, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 372 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 426, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 368 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 426, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 372 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 422, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 372 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 424, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 368 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 427, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 368 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 422, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 368 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 423, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 368 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 424, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 369 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 421, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 369 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 426, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 369 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 427, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 373 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 428, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 374 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 429, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 375 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 430, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 376 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 431, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 377 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 432, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 378 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 433, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 379 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 434, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 380 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 439, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 380 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 435, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 381 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 436, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 381 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 437, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 381 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 438, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 381 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 439, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 382 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 439, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 383 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 439, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 384 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 439, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 385 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 439, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 380, and / or the light chain comprises the amino acid sequence of SEQ ID NO: 440, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 384 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 440, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 383 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 436, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 383 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 438, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 385 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 436, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 385 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 438, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 380, and / or the light chain comprises the amino acid sequence of SEQ ID NO: 441, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 380 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 436, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 380 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 437, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 380 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 438, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 381 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 435, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 381 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 440, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 381 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 441, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 386 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 442, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 387 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 431, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 388 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 447, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 388 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 443, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 389 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 444, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 389 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 445, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 389 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 446, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 390 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 447, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 389 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 447, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 391 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 447, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 392 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 447, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 393 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 447, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 394 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 447, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 390 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 448, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 388 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 448, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 391 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 448, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 392 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 448, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 393 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 448, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 394 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 448, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 392 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 444, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 392 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 446, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 394 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 444, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 394 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 446, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 395 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 448, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 396 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 448, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 388 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 449, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 394 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 450, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 394 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 451, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 395 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 450, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 396 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 451, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 395 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 451, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 396 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 450, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 397 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 450, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 397 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 451, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 395 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 452, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 396 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 452, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 388 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 444, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 397 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 452, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 397 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 448, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 394 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 452, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 398 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 444, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 399 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 444, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 400 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 444, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 398 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 453, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 399 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 453, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 400 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 453, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 398 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 454, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 388 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 445, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 399 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 454, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 400 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 454, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 398 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 455, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 399 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 455, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 400 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 455, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 401 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 448, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 402 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 448, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 403 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 448, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 401 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 451, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 402 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 451, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 388 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 446, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 403 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 451, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 401 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 450, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 402 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 450, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 403 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 450, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 401 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 452, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 402 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 452, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 403 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 452, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 389 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 453, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 389 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 454, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 389 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 455, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 389 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 443, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 391 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 451, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 391 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 450, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 391 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 452, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 389 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 448, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 389 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 449, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 463 and / or the light chain comprises the amino acid sequence of SEQ ID NO: 464, or the heavy chain comprises the amino acid sequence of SEQ ID NO: 460, and / or the light chain comprises the amino acid sequence of SEQ ID NO: 404; The antibody or antigen-binding fragment thereof described in claim 7.

9. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof described in claim 1 and a pharmaceutically acceptable carrier.

10. An antibody or antigen-binding fragment thereof described in claim 1, or a pharmaceutical composition described in claim 9, for use in a method for treating congenital adrenal hyperplasia (CAH).

11. An antibody or antigen-binding fragment thereof described in claim 1, or a pharmaceutical composition described in claim 9, for use in a method for reducing androgen levels in a subject in need thereof.

12. An antibody or antigen-binding fragment thereof described in claim 1, or a pharmaceutical composition described in claim 9, for use in a method for reducing the severity of one or more symptoms or signs in a subject with CAH.

13. An antibody or antigen-binding fragment thereof described in claim 1, or a pharmaceutical composition described in claim 9, for use in a method for inhibiting the hypothalamic-pituitary-adrenal (HPA) axis in a subject in need of inhibition of the HPA axis.

14. An antibody or antigen-binding fragment thereof, or pharmaceutical composition for use according to any one of claims 10 to 13, wherein the method further comprises administering to the subject a mineralocorticoid and / or a glucocorticoid.

15. The antibody or antigen-binding fragment thereof, or pharmaceutical composition for use according to claim 10 or 12, wherein the CAH is associated with a mutation or deletion in the 21-hydroxylase gene (CYP21A2), or a mutation or deletion in the 11-beta-hydroxylase gene (CYP11B1), or a mutation or deletion in the 3-beta-hydroxysteroid dehydrogenase gene (HSD3B2).

Images