C-type natriuretic peptide grafted antibody

Abstract

The present invention relates to an antibody or a fragment thereof, which comprises at least one heterologous amino acid sequence incorporated into at least one CDR region of the antibody or its fragment, wherein the at least one heterologous amino acid sequence comprises an N-terminal linker sequence (Ntls), a C-type natriuretic peptide (CNP) and a C-terminal linker sequence (Ctls). Optionally, at least a portion of the at least one CDR region is replaced by the at least one heterologous amino acid sequence incorporated therein. The present invention also relates to such an antibody or its fragment for use in a method of treatment, a composition comprising such an antibody or its fragment, a nucleic acid or a mixture of nucleic acids encoding such an antibody or its fragment, a host cell comprising such a nucleic acid or such a mixture of nucleic acids, and a method for producing such an antibody or its fragment.

Description

Technical Field

[0001] The present invention relates to an antibody or fragment thereof, comprising at least one heterologous amino acid sequence incorporated into at least one CDR region of the antibody or fragment thereof, wherein the at least one heterologous amino acid sequence comprises an N-terminal linker sequence (Ntls), a C-type natriuretic peptide (CNP), and a C-terminal linker sequence (Ctls). Optionally, at least a portion of the at least one CDR region is replaced by the at least one heterologous amino acid sequence incorporated therein. There are at least 12 amino acid residues between: between amino acid residue HC (heavy chain) res25 according to Kabat and the first amino acid residue of CNP when the heterologous amino acid sequence is included in CDRH1; between amino acid residue HC res51 according to Kabat and the first amino acid residue of CNP when the heterologous amino acid sequence is included in CDRH2; between residue HC res92 according to Kabat and the first amino acid residue of CNP when the heterologous amino acid sequence is included in CDRH3; between residue LC (light chain) res26 according to Kabat and the first amino acid residue of CNP when the heterologous amino acid sequence is included in CDRL1; between amino acid residue LC res49 according to Kabat and the first amino acid residue of CNP when the heterologous amino acid sequence is included in CDRL2; and / or between amino acid residue LC res88 according to Kabat and the first amino acid residue of CNP when the heterologous amino acid sequence is included in CDRL3. In addition, there are at least 9 amino acid residues between the last amino acid residue of CNP and the following: amino acid residue HC res35a according to Kabat in case the heterologous amino acid sequence is included in CDRH1; amino acid residue HC res57 according to Kabat in case the heterologous amino acid sequence is included in CDRH2; amino acid residue HC res106 according to Kabat in case the heterologous amino acid sequence is included in CDRH3; amino acid residue LC res 32 according to Kabat in case the heterologous amino acid sequence is included in CDRL1; amino acid residue LC res57 according to Kabat in case the heterologous amino acid sequence is included in CDRL2; and / or amino acid residue LC res98 according to Kabat in case the heterologous amino acid sequence is included in CDRL3. The present invention also relates to the use of such an antibody or fragment thereof in a method of treatment, a composition comprising such an antibody or fragment thereof, a nucleic acid or a mixture of nucleic acids encoding such an antibody or fragment thereof, a host cell comprising such a nucleic acid or such a mixture of nucleic acids, and a method of producing such an antibody or fragment thereof. Background Art

[0002] Natriuretic peptides are a family of three structurally related peptides with neurohumoral effects. Atrial natriuretic peptide (ANP) is a 28-amino acid peptide containing a central ring structure formed by disulfide bridges between cysteine ​​residues 7 and 23. Human ANP is expressed in atrial myocytes as a 153-amino acid preprohormone. Signal peptide cleavage produces a prohormone form, which is then further cleaved into mature ANP and an N-terminal residue, known as NT-proANP. Similar to ANP, brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP) are also produced from precursor proteins and contain a central ring structure. ANP is primarily produced and released by the cardiomyocytes of the left and right atria, while BNP is primarily produced by ventricular cardiomyocytes. CNP is synthesized by vascular endothelial cells. In addition to these locations, smaller amounts of natriuretic peptides are also produced in other parts of the body, such as in the brain, kidneys, and adrenal glands. Natriuretic peptides are encoded by three independent genes: NPPA, NPPB, and NPPC. The amino acid sequences of these three peptides are highly conserved in mammals (Potter et al., Handb Exp Pharmacol. 2009; (191): 341-66). However, significant sequence modifications of natriuretic peptides have been documented, such as truncation, amino acid exchange, and chimeric fusions (e.g., CD-NP (McKie et al., Curr Heart Fail Rep. 2010 Sep; 7(3): 93-9)) to produce potent natriuretic peptides that activate or bind to cell receptors and can elicit relevant physiological effects.

[0003] Natriuretic peptides bind to three different membrane-bound receptor types, NPR-A, NPR-B, and NPR-C, mediating their biological effects. ANP and BNP bind with the greatest affinity to NPR-A; in contrast, CNP has the greatest affinity for the NPR-B receptor. NPR-A and NPR-B contain a (microparticle) guanylate cyclase domain (pGC), whose enzymatic activity leads to an increase in (intracellular) cyclic guanosine monophosphate (cGMP). As a second messenger, cGMP regulates various cellular processes. The NPR-C receptor does not exhibit guanylate cyclase activity and is also known as a "scavenger" receptor because it can bind natriuretic peptides, leading to their degradation by endocytosis. An additional signaling function for the NPR-C receptor has been described through regulation of cAMP (Anand-Srivastava, Peptides. 2005 Jun;26(6):1044-59).

[0004] The cardiac hormones ANP and BNP are excreted when the ventricles and atria are stretched, for example due to an excess of plasma volume. They exert a vasodilatory effect by relaxing vascular smooth muscle and leading to a decrease in blood pressure. In the kidneys, ANP leads to increased urine excretion (diuresis) and an increase in the concentration of sodium ions in the urine (natriuresis). ANP is considered a compensatory antagonist of the renin-angiotensin-aldosterone system (RAAS), which is overactivated in many cardiovascular diseases. In addition, ANP exerts other neurohumoral effects, including an inhibitory effect on the sympathetic nervous system and a complex regulatory effect on the baroreflex (Woods et al., Clin Exp Pharmacol Physiol. 2004 Nov; 31(11): 791-4). For ANP, as well as BNP and CNP, anti-inflammatory, anti-hypertrophic and anti-fibrotic effects have been demonstrated in animal models of different diseases (e.g., Knowles et al., 2001, J. Clin. Invest. 107: 975-984; Dahrouj et al., J Pharmacol Exp Ther. 2013 Jan; 344(1): 96-102; Baliga et al., Br J Pharmacol. 2014 Jul; 171(14): 3463-75; Mitaka et al. Intensive Care Med Exp. 2014 Dec; 2(1): 28; Werner et al., Basic Res Cardiol. 2016 Mar; 111(2): 22; Kimura et al., Respir Res. 2016 Feb 19; 17: 19). CNP activation of NPR-B plays an important role in bone growth (Yasoda et al., Clin. Calcium. 2009 Jul; 19(7): 1003-8) and vascular endothelial integrity (Moyes et al., J Clin Invest. 2014 Sep; 124(9): 4039-51).

[0005] The broad spectrum of physiological effects of natriuretic peptides and their receptors makes them attractive targets for drug development (Lumsden et al., Curr Pharm Des. 2010; 16(37): 4080-8; Buglioni et al., Annu Rev Med. 2016; 67: 229-43). For example, the natriuretic cGMP system may be inhibited in various pathophysiological conditions, which may lead to hypertension, increased cell proliferation, fibrosis, inflammation, endothelial dysfunction, diabetes, metabolic syndrome, atherosclerosis, cardiac insufficiency, myocardial infarction, pulmonary hypertension, eye and kidney diseases, bone disorders, stroke and / or sexual dysfunction.

[0006] The main obstacle to the therapeutic use of natriuretic peptides is that they have a very short plasma half-life in the body, which is only a few minutes (Hunt et al., J Clin Endocrinol Metab. 1994 Jun; 78(6): 1428-35; Kimura et al., Eur J Clin Pharmacol. 2007 Jul; 63(7): 699-702). In addition to endocytosis by NPR-C receptors, natriuretic peptides are also effectively proteolytically degraded by neprilysin (NEP) and insulin-degrading enzyme (IDE). The short-term biological effects of the administered natriuretic peptides limit their therapeutic use to acute indications. For example, in different countries, infusions of recombinant caperitide (ANP) and nesiritide (BNP) are approved for the treatment of acute decompensated heart failure.

[0007] The treatment of chronic diseases would be greatly advanced by providing NPR-A and NPR-B agonists with increased plasma half-life, improved proteolytic stability, and prolonged duration of action.

[0008] In recent years, several natriuretic peptide derivatives and variants have been described, such as CD-NP (McKie et al., Curr Heart Fail Rep. 2010 Sep; 7(3): 93-9), ZD100 / MANP (McKie et al., Hypertension. 2010 Dec; 56(6): 1152-9), PL-3994 (Edelson et al., Pulm Pharmacol Ther. 2013 Apr; 26(2): 229-38), ularitide (Anker et al., Eur Heart J. 2015 Mar 21; 36(12): 715-2), ANX-042 (Pan et al., Proc Natl Acad Sci USA. 2009 Jul. 7; 106(27): 11282-7), and BMN-111 (Wendt et al., J. Pharmacol Exp Ther. 2015 Apr; 353(1): 132-49. The half-life of CD-NP is approximately 18.5 minutes (Lee et al., BMC Pharmacology 2007, 7(Suppl 1): P38). Other ANP and CNP derivatives are disclosed in US 9,193,777 and EP 2432489 A, respectively.

[0009] In addition, natriuretic peptide fusions (including Fc fusions, albumin fusions) and pegylated natriuretic peptides are also described. For example, natriuretic peptide-Fc fusions are disclosed in US2010 / 0310561, WO2008 / 154226, WO2010 / 117760, WO2006 / 107124, WO2008 / 136611, and WO2008 / 079995. Natriuretic peptide-albumin fusions are disclosed in US7,521,424 and US2014 / 0148390, and pegylated natriuretic peptides are disclosed in US2014 / 0148390.

[0010] WO2005 / 060642 describes the generation of an ANP and BNP peptide-grafted antibody library obtained by inserting ANP or BNP with two random flanking amino acids at both ends into the CDRH3 region of a human tetanus toxoid-specific antibody. Similarly, WO2005 / 082004 discloses the generation of an ANP-mimicking grafted antibody library obtained by replacing the entire original CDRH3 region of the 2G12 antibody with an ANP-mimicking peptide flanked by two random amino acid residues. Neither WO2005 / 060642 nor WO2005 / 082004 discloses any specific natriuretic peptide-grafted antibodies, let alone functionally characterizing these antibodies.

[0011] Purpose of the Invention

[0012] In view of the prior art, the object of the present invention is to provide novel natriuretic peptide receptor agonists having increased stability in serum compared to naturally occurring wild-type natriuretic peptides. Summary of the Invention

[0013] The above-mentioned objects are achieved by the teachings of the present independent claims. The inventors surprisingly discovered that biologically active natriuretic peptide variants with significantly increased stability in serum compared to naturally occurring wild-type natriuretic peptides can be obtained by incorporating natriuretic peptide amino acid sequences into a CDR region of an immunoglobulin molecule or fragment thereof, despite the short length and high sequence conservation of the immunoglobulin CDR region, which has a considerable conformational inhibitory effect on the incorporation of the biologically active peptide. However, the activity of the natriuretic peptide incorporated into the immunoglobulin CDR region shows great variation. The inventors have found that the decisive factor for the successful incorporation of biologically active natriuretic peptide variants is the number of amino acid residues between the incorporated natriuretic peptide and the nearest neighboring CDR framework connecting the incorporated natriuretic peptide at the N-terminus and C-terminus. When the number of amino acid residues flanking the N-terminus and C-terminus between the natriuretic peptide and the neighboring CDR framework is less than a certain number, only natriuretic peptide-immunoglobulin fusion constructs with no biological activity or greatly reduced biological activity are obtained. Specific linker sequences flanking the incorporated natriuretic peptide have been found to be particularly advantageous for achieving high peptide activity, good expression levels and / or low protein fragmentation levels.

[0014] Thus, in a first aspect, the present invention relates to an antibody or fragment thereof comprising at least one heterologous amino acid sequence incorporated into at least one CDR region of the antibody or fragment thereof, wherein the at least one heterologous amino acid sequence comprises an N-terminal linker sequence (Ntls), a natriuretic peptide and a C-terminal linker sequence (Ctls), wherein optionally at least a portion of the at least one CDR region is replaced by the heterologous amino acid sequence incorporated therein,

[0015] a) at least 12 amino acid residues are present in

[0016] i) in case said heterologous amino acid sequence is incorporated into CDRH1, between amino acid residue HCres25 according to Kabat (which corresponds to res S25 in the heavy chain having the amino acid sequence SEQ ID NO 65) and the first amino acid residue of the natriuretic peptide;

[0017] ii) in case said heterologous amino acid sequence is incorporated into CDRH2, between amino acid residue HCres51 according to Kabat (which in the heavy chain having the amino acid sequence of SEQ ID NO 65 corresponds to res I51) and the first amino acid residue of the natriuretic peptide;

[0018] iii) in case said heterologous amino acid sequence is incorporated into CDRH3, between amino acid residue HCres92 according to Kabat (which corresponds to res C96 in the heavy chain having the amino acid sequence of SEQ ID NO 65) and the first amino acid residue of the natriuretic peptide;

[0019] iv) in case said heterologous amino acid sequence is incorporated into CDRL1, between amino acid residue LCres26 according to Kabat (which corresponds to res S25 in the light chain having the amino acid sequence SEQ ID NO 66) and the first amino acid residue of the natriuretic peptide;

[0020] v) between amino acid residue LCres49 according to Kabat (which in the light chain having the amino acid sequence SEQ ID NO 66 corresponds to res Y51) and the first amino acid residue of the natriuretic peptide in case said heterologous amino acid sequence is incorporated into CDRL2; and / or

[0021] vi) between amino acid residue LCres88 according to Kabat (which corresponds to res C90 in the light chain having the amino acid sequence SEQ ID NO 66) and the first amino acid residue of the natriuretic peptide in case said heterologous amino acid sequence is incorporated into CDRL3; and wherein

[0022] b) There are at least 9 amino acid residues between the last amino acid residue of the natriuretic peptide and

[0023] i) amino acid residue HCres35a according to Kabat (which corresponds to res M34 in the heavy chain having the amino acid sequence SEQ ID NO 65), in case said heterologous amino acid sequence is incorporated into CDRH1;

[0024] ii) amino acid residue HCres57 according to Kabat (which corresponds to res T58 in the heavy chain having the amino acid sequence SEQ ID NO 65), in case said heterologous amino acid sequence is incorporated into CDRH2;

[0025] iii) amino acid residue HCres106 according to Kabat (which corresponds to res G111 in the heavy chain having the amino acid sequence SEQ ID NO 65), in case said heterologous amino acid sequence is incorporated into CDRH3;

[0026] iv) amino acid residue LCres 32 according to Kabat (which corresponds to res D34 in the light chain having the amino acid sequence SEQ ID NO 66), in case said heterologous amino acid sequence is incorporated into CDRL1;

[0027] v) amino acid residue LCres57 according to Kabat (which corresponds to res G59 in the light chain having the amino acid sequence SEQ ID NO 66), in case said heterologous amino acid sequence is incorporated into CDRL2; and / or

[0028] vi) amino acid residue LCres98 according to Kabat (which corresponds to res F102 in the light chain having the amino acid sequence SEQ ID NO 66) in case said heterologous amino acid sequence is incorporated in CDRL3.

[0029] In other aspects, the present invention relates to the use of such antibodies or fragments thereof in methods of treatment, compositions comprising such antibodies or fragments thereof, nucleic acids or mixtures of nucleic acids encoding such antibodies or fragments thereof, host cells comprising such nucleic acids or such mixtures of nucleic acids, and methods of producing such antibodies or fragments thereof. DETAILED DESCRIPTION

[0030] The present invention may be understood more readily by reference to the following detailed description of the invention and the Examples included therein.

[0031] In a first aspect, the present invention relates to an antibody or fragment thereof, comprising at least one heterologous amino acid sequence incorporated into at least one CDR region of the antibody or fragment thereof, wherein the at least one heterologous amino acid sequence comprises an N-terminal linker sequence (Ntls), a natriuretic peptide and a C-terminal linker sequence (Ctls), wherein optionally a portion of the at least one CDR region is replaced by at least one heterologous amino acid sequence incorporated therein.

[0032] The present inventors have discovered that biologically active natriuretic peptide variants with significantly increased stability in serum compared to naturally occurring wild-type natriuretic peptides can be obtained by incorporating natriuretic peptide amino acid sequences into a CDR region of an immunoglobulin molecule or fragment thereof. This discovery was completely unexpected. As is well known in the art, the shorter length and higher sequence conservation of immunoglobulin CDR regions (particularly evident in CDRL1, CDRL2, CDRL3, CDRH1, and CDRH2) have considerable conformational inhibition on the incorporation of biologically active peptides, and the surrounding immunoglobulin sequences may adversely affect the expression, folding, and / or biological activity of the incorporated peptides. In fact, the present inventors have found that the activity of natriuretic peptides incorporated into immunoglobulin CDR regions varies greatly, depending on the exact method of constructing the natriuretic peptide-grafted antibody. The decisive factor for the successful incorporation of functional (i.e., biologically active) natriuretic peptide variants has been shown to be the number of amino acid residues between the N-terminus and the CDR framework connecting the incorporated natriuretic peptide to its nearest neighbor. When the N-terminal and C-terminal flanking amino acid residues between the natriuretic peptide and the adjacent CDR framework junction are below a certain number, no biologically active natriuretic peptide-immunoglobulin fusion construct is obtained.

[0033] The terms "incorporated," "inserted," "integrated," "grafted," and "embedded," as well as "incorporated," "inserted," "integrated," "grafted," and "embedded" are used interchangeably herein. In the context of the present invention, these terms refer to the generation of a hybrid polynucleic acid or hybrid polypeptide by the introduction of a heterologous sequence into the original sequence of an antibody or antibody fragment. Such incorporation can be performed in any manner. Typically, an antibody or fragment thereof comprising a natriuretic peptide flanked by N-terminal and C-terminal linker sequences is produced by recombinant DNA technology and expressed as described herein.

[0034] Incorporation of a natriuretic peptide flanked by N-terminal and C-terminal linker sequences into the CDR region of the original antibody or antibody fragment sequence may result in the deletion of at least a portion of the CDR region. For example, the nucleic acid sequence encoding the heterologous amino acid sequence comprising the N-terminal linker sequence, the natriuretic peptide, and the C-terminal linker sequence may be cloned such that a portion of the CDR encoding sequence is replaced by the incorporated heterologous nucleic acid sequence. In specific other embodiments, the incorporation of the heterologous amino acid sequence comprising the natriuretic peptide does not result in the deletion of amino acid residues in the CDR region into which the heterologous amino acid sequence is inserted.

[0035] In the context of the present invention, the term "heterologous amino acid sequence" refers to an amino acid sequence that is not derived from the original "empty" antibody or fragment thereof into which the amino acid sequence is incorporated. The heterologous amino acid sequence is grafted into the antibody or fragment thereof, thereby generating an engineered recombinant antibody molecule comprising amino acid sequences from a different source.

[0036] The term "natriuretic peptide" refers to a peptide that can induce increased natriuresis (i.e., sodium excretion by the kidneys). Natriuretic peptides include atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), C-type natriuretic peptide (CNP), Dendroaspis natriuretic peptide (DNP), and urodilatin. Natriuretic peptides within the meaning of the present invention may be of any origin. Natriuretic peptides include natural natriuretic peptides (such as wild-type natriuretic peptides) and mutant forms thereof, as well as homologous natriuretic peptides from different species. However, the term also includes modified natriuretic peptides, such as modified chimeric variants of different natriuretic peptides. It is known that codon usage varies between species. Therefore, when expressing heterologous proteins in target cells, it may be necessary or at least beneficial to modify the nucleic acid sequence to adapt to the codon usage of the target cells. Methods for designing and constructing derivatives of a given protein are well known to any person skilled in the art.

[0037] In a specific embodiment, the natriuretic peptide is selected from the wild-type natriuretic peptide of any species and a functional variant of any such wild-type natriuretic peptide. In the context of the present invention, the term "functional variant of a natriuretic peptide" or "functional natriuretic peptide variant" refers to a natriuretic peptide of any origin, including natural peptides and modified peptides, which differ in amino acid sequence and / or nucleic acid sequence encoding the amino acid sequence of a given natriuretic peptide (e.g., a wild-type natriuretic peptide of a given species), but still have functional activity. In the context of the present invention, the term "functional activity" refers to the ability of a natriuretic peptide variant to perform the biological function of a naturally occurring natriuretic peptide (particularly a wild-type natriuretic peptide). Specifically, "functional activity" means that the natriuretic peptide variant is able to bind to its respective receptors. In the case of NPR-A and NPR-B ligands, "functional activity" specifically means the ability to mediate an increase in (intracellular) cyclic guanosine monophosphate (cGMP) by binding to one or both of these receptors.

[0038] In specific embodiments, the functional natriuretic peptide variant is capable of performing at least about 50%, specifically at least about 60%, at least about 70%, at least about 80%, and most specifically at least about 90% of one or more biological functions of a given natriuretic peptide (e.g., a wild-type natriuretic peptide of any given species), wherein the one or more biological functions include but are not limited to binding of the natriuretic peptide to its respective receptor and / or inducing an increase in intracellular cGMP.

[0039] The functional activity of the natriuretic peptide can be measured by any method (including in vitro methods) that makes it possible to measure an increase in (intracellular) cyclic guanosine monophosphate (cGMP) or to measure changes in cGMP-regulated cellular processes, including the methods described in Examples 3 and 5. In a specific embodiment, if the EC of the (non-grafted) natriuretic peptide variant determined by the fluorometric assay described in Example 3 is 50A value of less than 500 nM, more specifically less than 250 nM, more specifically less than 150 nM, more specifically less than 100 nM, more specifically less than 50 nM, most specifically less than 25 nM is considered to be functionally active.

[0040] This functional natriuretic peptide variant is incorporated into a CDR region of an immunoglobulin molecule or fragment thereof as described herein to produce a natriuretic peptide grafted immunoglobulin having natriuretic peptide functional activity, and the stability of the natriuretic peptide grafted immunoglobulin in serum is significantly increased compared to the non-grafted functional natriuretic peptide variant, as shown in the Examples. If the natriuretic peptide grafted immunoglobulin gives a significant positive signal in any method that directly or indirectly measures the increase in (intracellular) cyclic guanosine monophosphate (cGMP) by evaluating changes in cGMP-regulated cellular processes, the natriuretic peptide grafted immunoglobulin is considered to be biologically active (i.e., functional). Specifically, the functional activity of the natriuretic peptide grafted immunoglobulin can be evaluated by the methods described in Examples 3 and 5. In the case of natriuretic peptide grafted immunoglobulins, significance is typically assessed based on: i) comparison to negative samples, such as an empty immunoglobulin scaffold, e.g., construct #209 (antibody comprising SEQ ID NO 65 and SEQ ID NO 66, TPP-5657), ii) comparison to positive samples, e.g., construct #117 (antibody comprising SEQ ID NO 67 and SEQ ID NO 66, TPP-5661), and iii) dose dependency.

[0041] Although the functional natriuretic peptide variants of the present invention may contain any number of mutations comprising additions, deletions and / or substitutions of one or more amino acids compared to the reference natriuretic peptide, the functional natriuretic peptide variants will generally retain the key features of the corresponding natriuretic peptide, such as key residues within the central loop domain. For example, the conserved residues of natriuretic peptides are described in Lincoln R. Potter et al. (Handb Exp Pharmacol. 2009; (191): 341-366). Thus, in specific embodiments, the functional natriuretic peptide variants have at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% sequence identity with the sequence shown below:

[0042] X1CFGX2X3X4DRIX5X6X7SX8LGC

[0043] Where X1 and X5 are G or S;

[0044] X3 is R or K;

[0045] X6 is A or S; and

[0046] X2, X4, X7 and X8 can be any amino acid.

[0047] In principle, any type of natriuretic peptide can be incorporated into the CDR region of an immunoglobulin or fragment thereof as described herein. Specifically, the inventors have found that the minimum requirements for satisfactory biological activity of the grafted natriuretic peptide and the discovery of particularly suitable N-terminal and C-terminal amino acid sequences in one type of natriuretic peptide can be imparted to other types of natriuretic peptides. Without wishing to be bound by theory, it is speculated that these similar requirements for successfully embedding natriuretic peptides into immunoglobulin molecules in different types of natriuretic peptides may be due to structural similarities and / or mechanisms of action within the natriuretic peptide family.

[0048] In a specific embodiment, the natriuretic peptide is selected from the group consisting of human ANP having the sequence of SEQ ID NO 23, human BNP having the sequence of SEQ ID NO 24, human CNP having the sequence of SEQ ID NO 25, and peptides having at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or at least 98% sequence identity to any one of SEQ ID NOs 23 to 25. Similarly, natriuretic peptides having sequences that deviate from the wild-type human natriuretic peptides ANP, BNP and CNP may be of any natural origin, such as mutant versions of wild-type human natriuretic peptides or homologs from different species or engineered natriuretic peptides. Methods for designing and constructing peptide variants are known to any one of ordinary skill in the art.

[0049] In specific such embodiments, a natriuretic peptide having at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or at least 98% sequence identity to any one of SEQ ID NOs 23 to 25 is a functional natriuretic peptide variant.

[0050] For reference polynucleotide or polypeptide sequences, "percent (%) sequence identity" is defined as the percentage of nucleic acid or amino acid residues in a candidate sequence that are identical to the nucleic acid or amino acid residues in a reference polynucleotide or polypeptide sequence, respectively, after alignment of the sequences and, if necessary, the optionally introduced gaps to achieve maximum percent sequence identity. Conservative substitutions are not considered part of sequence identity. In specific embodiments, the total amount of any gaps introduced into a candidate sequence and / or reference sequence does not exceed 50%, 40%, 30%, 25%, 20%, 15% or 10% of the total amount of residues in the reference sequence. In specific embodiments, percent sequence identity is determined without introducing any gaps into the candidate sequence or reference sequence (i.e., using a gapless alignment). Alignment for determining percent amino acid sequence identity can be achieved in various ways fully understood by those skilled in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithm required for achieving maximum alignment over the full length of the compared sequences.

[0051] A natriuretic peptide having a given percentage of sequence identity with a given reference natriuretic peptide (e.g., human CNP having the amino acid sequence of SEQ ID NO 25) may contain one or more mutations comprising the addition, deletion, and / or substitution of one or more amino acids compared to the reference natriuretic peptide. According to the teachings of the present invention, the amino acids deleted, added, and / or substituted may be continuous amino acids, or may be dispersed over the length of the amino acid sequence of a natriuretic peptide having a given percentage of sequence identity with a reference natriuretic peptide (e.g., human CNP having the amino acid sequence of SEQ ID NO 25). At the DNA level, nucleic acid sequences encoding natriuretic peptides having a given percentage of sequence identity with a given reference natriuretic peptide may differ to a greater extent due to the degeneracy of the genetic code.

[0052] According to the teachings of the present invention, any number of amino acids may be added, deleted and / or substituted as long as the specified amino acid sequence identity with the reference natriuretic peptide is maintained. In a specific embodiment, the specified amino acid sequence identity is maintained and the natriuretic peptide variant has biological activity, i.e., it is a functional natriuretic peptide variant. Preferably, the reduction in biological activity of a natriuretic peptide having a given sequence identity percentage with a given reference natriuretic peptide (e.g., human CNP having the amino acid sequence of SEQ ID NO 25) is less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 25% or less than 10% compared to the reference natriuretic peptide measured in the above-mentioned assay.

[0053] As used herein, the term "antibody" refers to an immunoglobulin molecule, particularly a dimeric immunoglobulin molecule composed of four polypeptide chains: two heavy (H) chains and two light (L) chains, which are typically interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region can include, for example, three domains: CH1, CH2, and CH3. Each light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region consists of one domain (CL). The VH and VL regions can be further subdivided into highly variable regions, called complementarity determining regions (CDRs), with more conserved regions in between, called framework regions (FRs). Each VH and VL typically consists of three CDRs and four FRs arranged from amino-terminus to carboxyl-terminus, for example in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0054] As used herein, the term "complementarity determining region" (CDR; e.g., CDR1, CDR2, and CDR3) refers to the amino acid residues of an antibody variable domain whose presence is necessary for antigen binding. Each variable domain typically has three CDR regions, identified as CDR1, CDR2, and CDR3. Each complementarity determining region can include amino acid residues in a "complementarity determining region" as defined by Kabat (e.g., about residues 24-34 (CDRL1), 50-56 (CDRL2), and 89-97 (CDRL3) in a light chain variable domain, and 31-35 (CDRH1), 50-65 (CDRH2), and 95-102 (CDRH3) in a heavy chain variable domain; (Kabat et al., Sequences of Proteins of Immulological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)) and / or those residues from the "hypervariable loops" (e.g., approximately residues 26-32 (CDRL1), 50-52 (CDRL2), and 91-96 (CDRL3) in the light chain variable domain, and residues 26-32 (CDRH1), 53-55 (CDRH2), and 96-101 (CDRH3) in the heavy chain variable domain (Chothia and Lesk; J Mol Biol 196:901-917 (1987)). In certain cases, the complementarity determining regions may include amino acids from both CDR regions and hypervariable loops according to the Kabat definition.

[0055] According to the amino acid sequence of its heavy chain constant domain, complete antibodies can be assigned to different "classes". There are five major classes of complete antibodies: IgA, IgD, IgE, IgG and IgM, several of which can be further divided into "subclasses" (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. In the context of the present invention, the term "antibody" includes any main class of immunoglobulin molecules (including IgG, IgE, IgM, IgD, IgA and IgY) and any subclass of immunoglobulin molecules (including IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) isolated from nature or prepared using recombinant methods, and includes all conventionally known antibodies. A preferred class of immunoglobulins for use in the present invention is IgG. The term "antibody" also extends to other protein scaffolds that are capable of orienting antibody CDR inserts into an active binding conformation that is identical to that present in a native antibody, such that the observed binding of the target antigen to these chimeric proteins is unchanged compared to the binding activity of the native antibody from which the CDRs were derived.

[0056] In the context of the present invention, the term "fragment" of an antibody / immunoglobulin refers to any portion of an antibody / immunoglobulin that comprises at least one CDR region. In particular, the antibody fragments of the present invention retain the ability to increase the serum half-life of a biologically active peptide (preferably a natriuretic peptide) incorporated therein. The antibody fragments of the present invention include Fab, Fab', Fab'-SH, F(ab')2 and Fv fragments; diabodies; single domain antibodies (Dab); linear antibodies; single-chain antibody molecules (scFv); and disulfide-stabilized Fv antibody fragments (dsFv); as well as multispecific antibodies formed from antibody fragments and fragments comprising VL or VH domains, which are prepared from intact immunoglobulins or by recombinant methods.

[0057] F(ab')2 or Fab can be engineered to minimize or completely eliminate the intermolecular disulfide interactions between the CH1 and CL domains. Antibody fragments of the present invention can include the variable region alone or in combination with all or part of the hinge region, CH1, CH2, CH3, and CL domains. Also included are antibody fragments comprising any combination of the variable region and the hinge region, CH1, CH2, CH3, and CL domains.

[0058] The antibodies or fragments thereof constitute a scaffold that confers stability to the natriuretic peptide incorporated therein. For example, the serum half-life of a natriuretic peptide incorporated into the CDR regions of the antibodies described herein may be increased compared to a naturally occurring natriuretic peptide.

[0059] Primarily, the heterologous amino acid sequence comprising a natriuretic peptide can be incorporated into any immunoglobulin molecule or its fragment. Specifically, any species (including but not limited to humans, cattle, mice, rats, pigs, dogs, sharks, llamas (lama) and camels) and any main class and subclass immunoglobulins can be used according to the present invention. However, for therapeutic use, humans or humanized antibodies may be preferred. In the context of the present invention, the term "human antibody" refers to an antibody with a human immunoglobulin amino acid sequence, including antibodies isolated from a human immunoglobulin library, from human B cells, or from animals of transgenic one or more human immunoglobulins, as well as synthetic human antibodies. In a specific embodiment, the amino acid light chain and heavy chain sequences of the variable domains are derived from human germline sequences LV 1-40 and HV 3-23, respectively (for more information, see Example 1).

[0060] In the context of the present invention, the term "humanized antibody" or "humanized antibody fragment" refers to (i) an antibody or fragment thereof derived from a non-human source (e.g., a transgenic mouse carrying a heterologous immune system), which antibody is based on human germline sequences; or (ii) a chimeric antibody or fragment thereof, in which the variable domain is derived from a non-human source and the constant domain is derived from a human source, or (iii) a CDR-grafted antibody or fragment thereof, in which the CDRs of the variable domain are derived from a non-human source, while one or more frameworks of the variable domain are of human origin and the constant domain (if any) is of human origin.

[0061] The antibodies of the invention or fragments thereof may be monospecific, bispecific, trispecific or of greater multispecificity.

[0062] In the context of the present invention, the term "comprises" or "comprising" means "including but not limited to". The term is intended to be open-ended to specify the presence of any claimed feature, element, integer, step, or component, but does not preclude the presence or addition of one or more other features, elements, integers, steps, components, or groups thereof. Thus, the term "comprising" encompasses the more restrictive terms "consisting of" and "consisting essentially of". In one embodiment, the term "comprising" as used throughout the application and particularly in the claims may be replaced with the term "consisting of".

[0063] In the context of the present invention, the term "approximately" or "about" means between 80% and 120%, or between 90% and 110%, including between 95% and 105%, of a given value.

[0064] In the antibodies or fragments thereof of the present invention, a) at least 12 amino acid residues are present in

[0065] i) in case a heterologous amino acid sequence is incorporated into CDRH1, between amino acid residue HC res25 according to Kabat (which corresponds to res S25 in the heavy chain having the amino acid sequence SEQ ID NO 65) and the first amino acid residue of the natriuretic peptide;

[0066] ii) in case a heterologous amino acid sequence is incorporated into CDRH2, between amino acid residue HC res51 according to Kabat (which in the heavy chain having the amino acid sequence SEQ ID NO 65 corresponds to res I51) and the first amino acid residue of the natriuretic peptide;

[0067] iii) in case a heterologous amino acid sequence is incorporated into CDRH3, between amino acid residue HC res92 according to Kabat (which corresponds to res C96 in the heavy chain having the amino acid sequence of SEQ ID NO 65) and the first amino acid residue of the natriuretic peptide;

[0068] iv) in case a heterologous amino acid sequence is incorporated into CDRL1, between amino acid residue LC res26 according to Kabat (which corresponds to res S25 in the light chain having the amino acid sequence SEQ ID NO 66) and the first amino acid residue of the natriuretic peptide;

[0069] v) in case a heterologous amino acid sequence is incorporated into CDRL2, between amino acid residue LC res49 according to Kabat (which corresponds to res Y51 in the light chain having the amino acid sequence SEQ ID NO 66) and the first amino acid residue of the natriuretic peptide; and / or

[0070] vi) in case a heterologous amino acid sequence is incorporated into CDRL3, between amino acid residue LC res88 according to Kabat (which corresponds to res C90 in the light chain having the amino acid sequence SEQ ID NO 66) and the first amino acid residue of the natriuretic peptide;

[0071] and b) there are at least 9 amino acid residues between the last amino acid residue of the natriuretic peptide and

[0072] i) amino acid residue HC res35a according to Kabat (which corresponds to res M34 in the heavy chain having the amino acid sequence SEQ ID NO 65), in case a heterologous amino acid sequence is incorporated into CDRH1;

[0073] ii) amino acid residue HC res57 according to Kabat (which corresponds to res T58 in the heavy chain having the amino acid sequence SEQ ID NO 65) in case a heterologous amino acid sequence is incorporated into CDRH2;

[0074] iii) in case a heterologous amino acid sequence is incorporated into CDRH3, amino acid residue HCres106 according to Kabat (which corresponds to res G111 in the heavy chain having the amino acid sequence SEQ ID NO 65);

[0075] iv) amino acid residue LC res 32 according to Kabat (in the light chain having the amino acid sequence SEQ ID NO 66, which corresponds to res D34) in case a heterologous amino acid sequence is incorporated into CDRL1;

[0076] v) amino acid residue LCres57 according to Kabat (which corresponds to resG59 in the light chain having the amino acid sequence SEQ ID NO 66) in case a heterologous amino acid sequence is incorporated into CDRL2; and / or

[0077] vi) In case of inclusion of a heterologous amino acid sequence in CDRL3, amino acid residue LC res98 according to Kabat (in the light chain having the amino acid sequence SEQ ID NO 66, which corresponds to res F102).

[0078] The nomenclature of the amino acid residues listed above refers to the amino acid position in the original immunoglobulin molecule before the inclusion of the heterologous amino acid sequence. In the context of the present invention, the amino acid residues listed above are referred to as "reference amino acids" or "reference aa". These reference amino acid residues are located at or near the CDR framework junctions, but do not necessarily correspond to the standard CDR boundary definitions (standard CDR boundary definitions are amino acid residues S25 and W36 of CDRH1; S49 and R67 of CDRH2; K98 and W108 of CDRH3; C22 and W37 of CDRL1; Y51 and G59 of CDRL2; C90 and F102 of CDRL3. Jarasch and Skerra, Proteins 2017 Jan; 85(1): 65-71).

[0079] The N-terminal nearest reference aa of the inserted natriuretic peptide plus the amino acid stretch between the reference aa and the first amino acid residue of the inserted natriuretic peptide is referred to herein as the "N-terminal sequence". The N-terminal sequence comprises Ntls. In a specific embodiment, the N-terminal sequence consists of Ntls plus the adjacent N-terminal reference aa.

[0080] The amino acid stretch between the last amino acid residue of the inserted natriuretic peptide and the reference aa nearest to the C-terminus of the inserted natriuretic peptide plus the reference aa is referred to herein as the "C-terminal sequence". The C-terminal sequence comprises Ctls. In a specific embodiment, the C-terminal sequence consists of Ctls plus the adjacent C-terminal reference aa.

[0081] In a specific embodiment, Ntls comprises a GS linker sequence; a PN linker sequence; an amino acid sequence that is part of a human IgG antibody scaffold, or a sequence having at least 80% sequence identity thereto, particularly an amino acid sequence that is part of a fab domain scaffold of a human IgG antibody, or a sequence having at least 80% sequence identity thereto, more particularly a sequence of any one of SEQ ID NOs 1, 2, or 4, or a sequence having at least 80% sequence identity thereto; a sequence of any one of SEQ ID NOs 6, 7, 9, 11, 13, 15, 16, 17, 19, or 21; or a sequence having at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% sequence identity thereto. Ntls may also comprise any combination of the amino acid sequences listed above.

[0082] In specific such embodiments, the Ntls comprises a GS linker sequence; a PN linker sequence; an amino acid sequence that is part of a human IgG antibody scaffold, or a sequence having at least 80% sequence identity thereto, in particular an amino acid sequence that is part of a fab domain scaffold of a human IgG antibody, or a sequence having at least 80% sequence identity thereto, more particularly a sequence of any one of SEQ ID NOs 1, 2 or 4, or a sequence having at least 80% sequence identity thereto; a sequence of any one of SEQ ID NOs 6, 7, 9, 11, 13, 15 or 21; a sequence having at least 60%, at least 70%, at least 80%, at least 90% or at least 95% sequence identity thereto; or any combination thereof.

[0083] In a specific embodiment, Ctls comprises a GS linker sequence; a PN linker sequence; an amino acid sequence that is part of a human IgG antibody scaffold, or a sequence having at least 80% sequence identity thereto, particularly an amino acid sequence that is part of a fab domain scaffold of a human IgG antibody, or a sequence having at least 80% sequence identity thereto, more particularly a sequence of any one of SEQ ID NOs 1, 3, or 5, or a sequence having at least 80% sequence identity thereto; a sequence of any one of SEQ ID NOs 6, 8, 10, 12, 14, 15, 17, 18, 19, 20, or 22; or a sequence having at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% sequence identity thereto. Ctls may also comprise any combination of the amino acid sequences listed above.

[0084] In a specific embodiment, Ctls comprises a GS linker sequence; a PN linker sequence; an amino acid sequence that is part of a human IgG antibody scaffold, or a sequence having at least 80% sequence identity thereto, in particular an amino acid sequence that is part of a fab domain scaffold of a human IgG antibody, or a sequence having at least 80% sequence identity thereto, more particularly a sequence of any one of SEQ ID NOs 1, 3 or 5, or a sequence having at least 80% sequence identity thereto; a sequence of any one of SEQ ID NOs 6, 8, 10, 12, 14, 15, 20 or 22; a sequence having at least 60%, at least 70%, at least 80%, at least 90% or at least 95% sequence identity thereto; or any combination thereof.

[0085] In a specific embodiment, the sequence contained in Ntls and / or Ctls has a sequence identity of at least 60%, specifically at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% with any one of SEQ ID NOs 1 to 22.

[0086] In the context of the present invention, the term "GS linker sequence" refers to a peptide linker that primarily comprises glycine and serine residues. Specifically, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% of the amino acid residues of the GS linker sequence of the present invention are selected from glycine and serine residues. The GS linker sequence of the present invention may, for example, comprise a total of 1 to 30 amino acid residues. Specifically, the GS linker sequence of the present invention does not comprise more than 3, 2, or 1 amino acid residues that are not glycine or serine.

[0087] In the context of the present invention, the term "PN linker sequence" refers to a peptide linker that primarily comprises proline and asparagine residues. Specifically, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% of the amino acid residues of the PN linker sequence of the present invention are selected from proline and asparagine residues. The PN linker sequence of the present invention may, for example, comprise a total of 1 to 30 amino acid residues. Specifically, the PN linker sequence of the present invention does not comprise more than 3, 2, or 1 amino acid residues that are not proline or asparagine. Other amino acid residues that may be present in the PN linker sequence of the present invention are, for example, lysine or glutamic acid residues.

[0088] In a specific embodiment, the linker sequence contained within Ntls and / or Ctls and selected from the group consisting of: a GS linker sequence; a PN linker sequence; a human IgG antibody scaffold linker sequence; a human IgG Fab domain scaffold sequence; a sequence having at least 80% sequence identity to a human IgG antibody scaffold linker sequence, a human IgG Fab domain scaffold sequence, or a sequence of any one of SEQ ID NOs 1 to 5; or a sequence having at least 60% sequence identity to any one of SEQ ID NOs 6 to 22. The linker sequence contained within Ntls and / or Ctls may, for example, comprise up to 30, 28, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, or 10 amino acid residues.

[0089] In the case of a linker comprising a sequence of a human IgG antibody scaffold or a sequence having at least 80% sequence identity thereto, it may be particularly advantageous to use a sequence from the scaffold region adjacent to the CDR incorporating the heterologous amino acid sequence. For example, in the case of a heterologous amino acid sequence incorporating the CDRH2 domain, Ntls and / or Ctls may comprise a linker comprising an amino acid sequence that is part of the framework region FRH2 or FRH3. Similarly, in the case of a heterologous amino acid sequence incorporating the CDRL2 region, Ntls and / or Ctls may comprise a linker comprising an amino acid sequence that is part of the framework region FRH2 or FRH3.

[0090] In a specific embodiment, Ntls consists of the following: a GS linker sequence; a PN linker sequence; an amino acid sequence that is part of a human IgG antibody scaffold, or a sequence having at least 80% sequence identity thereto, in particular an amino acid sequence that is part of a fab domain scaffold of a human IgG antibody, or a sequence having at least 80% sequence identity thereto, more particularly a sequence of any one of SEQ ID NOs 1, 2 or 4, or a sequence having at least 80% sequence identity thereto; a sequence of any one of SEQ ID NOs 6, 7, 9, 11, 13, 15, 16, 17, 19 or 21; a sequence having at least 60%, at least 70%, at least 80%, at least 90% or at least 95% sequence identity thereto; or any combination thereof.

[0091] In a specific embodiment, Ctls consists of the following: a GS linker sequence; a PN linker sequence; an amino acid sequence that is part of a human IgG antibody scaffold, or a sequence having at least 80% sequence identity thereto, in particular an amino acid sequence that is part of a fab domain scaffold of a human IgG antibody, or a sequence having at least 80% sequence identity thereto, more in particular a sequence of any one of SEQ ID NOs 1, 3 or 5, or a sequence having at least 80% sequence identity thereto; a sequence of any one of SEQ ID NOs 6, 8, 10, 12, 14, 15, 17, 18, 19, 20 or 22; a sequence having at least 60%, at least 70%, at least 80%, at least 90% or at least 95% sequence identity thereto; or any combination thereof.

[0092] In specific embodiments, both Ntls and Ctls comprise at least one of the linker sequences listed above, or any combination thereof. In principle, any of the Ntls linker sequences listed above can be combined with any of the Ctls linker sequences listed above. Specifically, any linker sequence can be combined with a GS linker. As a non-limiting example, a GS Ctls linker can be combined with an Ntls linker comprising any of SEQ ID NOs: 6, 9, or 15, or a sequence having at least 60% sequence identity to any of SEQ ID NOs: 6, 9, or 15. Another non-limiting example is a GS Ntls linker combined with a Ctls linker comprising SEQ ID NO: 15, or a sequence having at least 60% sequence identity thereto.

[0093] In the case where the total length of the N-terminal and C-terminal flanking sequences is sufficient, the linker sequences listed above have been shown to be particularly advantageous for obtaining good natriuretic peptide activity. Without wishing to be bound by theory, it is believed that the linker peptide segments listed above result in a conformation / folding that promotes a system state that is favorable for presenting the biologically active natriuretic peptide to its respective receptor with minimal steric hindrance.

[0094] In specific embodiments, i) Ntls comprises a GS linker sequence; a PN linker sequence; a sequence of SEQ ID NO 2, 4, 9, 11, 13 or 15; a sequence having at least 60% sequence identity to SEQ ID NO 2, 4, 9, 11, 13 or 15; or any combination thereof, and ii) Ctls comprises a GS linker sequence; a PN linker sequence; a sequence of SEQ ID NO 3, 5, 12, 14, 15 or 20; a sequence having at least 60% sequence identity to SEQ ID NO 3, 5, 12, 14, 15 or 20; or any combination thereof. These linker sequences have been shown to be particularly useful because they not only achieve high natriuretic peptide activity, but also have good expression levels in recombinant expression and are less susceptible to protein fragmentation during expression (see Table 9).

[0095] In a specific embodiment, Ntls and Ctls each comprise a GS linker sequence; Ntls and Ctls each comprise a PN linker sequence; Ntls and Ctls each comprise an amino acid sequence that is part of a human IgG antibody scaffold, or a sequence having at least 80% sequence identity thereto, specifically an amino acid sequence that is part of a fab domain scaffold of a human IgG antibody, or a sequence having at least 80% sequence identity thereto, more specifically Ntls comprises the sequence of any one of SEQ ID NOs 1, 2 or 4, or a sequence having at least 80% sequence identity thereto, Ctls comprises the sequence of any one of SEQ ID NOs 1, 3 or 5, or a sequence having at least 80% sequence identity thereto; Ntls and Ctls each comprise the sequence of SEQ ID NO 6, or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 7, or a sequence having at least 60% sequence identity thereto, Ctls comprises the sequence of SEQ ID NO 8, or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 9 or a sequence having at least 60% sequence identity thereto, Ctls comprises the sequence of SEQ ID NO 10 or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 11 or a sequence having at least 60% sequence identity thereto, Ctls comprises the sequence of SEQ ID NO 12 or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 13 or a sequence having at least 60% sequence identity thereto, Ctls comprises the sequence of SEQ ID NO 14 or a sequence having at least 60% sequence identity thereto; Ntls and Ctls each comprise the sequence of SEQ ID NO 15 or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 16 or a sequence having at least 60% sequence identity thereto, Ctls comprises the sequence of SEQ ID NO 17 or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 18 or a sequence having at least 60% sequence identity thereto 17 or a sequence having at least 60% sequence identity thereto, Ctls comprises the sequence of SEQ ID NO 18 or a sequence having at least 60% sequence identity thereto; Ntls and Ctls each comprise the sequence of SEQ ID NO 19 or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 9 or a sequence having at least 60% sequence identity thereto, and Ctls comprises the sequence of SEQ ID NO 20 or a sequence having at least 60% sequence identity thereto;or Ntls comprises the sequence of SEQ ID NO 21 or a sequence having at least 60% sequence identity thereto, and Ctls comprises the sequence of SEQ ID NO 22 or a sequence having at least 60% sequence identity thereto. ;

[0096] In specific such embodiments, Ntls and Ctls each comprise a GS linker sequence; Ntls and Ctls each comprise a PN linker sequence; Ntls and Ctls each comprise an amino acid sequence that is part of a human IgG antibody scaffold or a sequence having at least 80% sequence identity thereto, specifically an amino acid sequence that is part of a fab domain scaffold of a human IgG antibody or a sequence having at least 80% sequence identity thereto, more specifically Ntls comprises a sequence of any one of SEQ ID NOs 1, 2 or 4 or a sequence having at least 80% sequence identity thereto, Ctls comprises a sequence of any one of SEQ ID NOs 1, 3 or 5 or a sequence having at least 80% sequence identity thereto; Ntls and Ctls each comprise the sequence of SEQ ID NO 6 or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 7 or a sequence having at least 60% sequence identity thereto, Ctls comprises the sequence of SEQ ID NO 8 or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 9 or a sequence with at least 60% sequence identity thereto, Ctls comprises the sequence of SEQ ID NO 10 or a sequence with at least 60% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 11 or a sequence with at least 60% sequence identity thereto, Ctls comprises the sequence of SEQ ID NO 12 or a sequence with at least 60% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 13 or a sequence with at least 60% sequence identity thereto, Ctls comprises the sequence of SEQ ID NO 14 or a sequence with at least 60% sequence identity thereto; Ntls and Ctls each comprise the sequence of SEQ ID NO 15 or a sequence with at least 60% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 9 or a sequence with at least 60% sequence identity thereto, Ctls comprises the sequence of SEQ ID NO 20 or a sequence with at least 60% sequence identity thereto; or Ntls comprises the sequence of SEQ ID NO 21 or a sequence with at least 60% sequence identity thereto, Ctls comprises the sequence of SEQ ID NO 22 or a sequence having at least 60% sequence identity thereto.

[0097] In specific such embodiments, Ntls and Ctls each comprise a GS linker sequence; Ntls and Ctls each comprise a PN linker sequence; Ntls comprises the sequence of SEQ ID NO 2 or a sequence having at least 80% sequence identity thereto, and Ctls comprises the sequence of SEQ ID NO 3 or a sequence having at least 80% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 4 or a sequence having at least 80% sequence identity thereto, and Ctls comprises the sequence of SEQ ID NO 5 or a sequence having at least 80% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 11 or a sequence having at least 60% sequence identity thereto, and Ctls comprises the sequence of SEQ ID NO 12 or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence of SEQ ID NO 13 or a sequence having at least 60% sequence identity thereto, and Ctls comprises the sequence of SEQ ID NO 14 or a sequence having at least 60% sequence identity thereto; Ntls and Ctls each comprise the sequence of SEQ ID NO 15 or a sequence having at least 60% sequence identity thereto; or Ntls comprises the sequence of SEQ ID NO 16 or a sequence having at least 60% sequence identity thereto. ID NO 9 or a sequence having at least 60% sequence identity thereto, and Ctls comprises the sequence SEQ ID NO 20 or a sequence having at least 60% sequence identity thereto. These linker combinations have been shown to be particularly useful for obtaining high natriuretic peptide activity, good expression levels in recombinant expression, and minimal or no protein fragmentation, as shown in Table 9.

[0098] In specific embodiments, Ntls further comprises an anchoring element A1 at its C-terminus and / or Ctls further comprises an anchoring element A2 at its N-terminus, wherein A1 and A2 primarily comprise glycine and serine residues. In specific embodiments, A1 and / or A2 comprise at least 1, 2, 3, 4, or 5 amino acid residues. A1 and / or A2 may comprise up to 10, 9, 8, 7, 6, or 5 amino acid residues in total. In specific embodiments, at least 60%, at least 70%, at least 80%, at least 90%, or 100% of the amino acid residues of A1 and / or A2 are selected from glycine and serine residues. Specifically, A1 and / or A2 do not comprise more than 3, 2, or 1 amino acid residues other than glycine or serine.

[0099] In a specific embodiment, Ntls consists of: i) an anchoring element A1 at its C-terminus and ii) a GS linker sequence; a PN linker sequence; an amino acid sequence that is part of a human IgG antibody scaffold or a sequence having at least 80% sequence identity thereto, in particular an amino acid sequence that is part of a fab domain scaffold of a human IgG antibody or a sequence having at least 80% sequence identity thereto, more particularly a sequence of any one of SEQ ID NOs 1, 2 or 4 or a sequence having at least 80% sequence identity thereto; a sequence of any one of SEQ ID NOs 6, 7, 9, 11, 13, 15, 16, 17, 19 or 21; a sequence having at least 60% sequence identity thereto; or any combination thereof.

[0100] In a specific embodiment, Ctls consists of: i) an anchoring element A2 at its N-terminus and ii) a GS linker sequence; a PN linker sequence; an amino acid sequence that is part of a human IgG antibody scaffold or a sequence having at least 80% sequence identity thereto, in particular an amino acid sequence that is part of a fab domain scaffold of a human IgG antibody or a sequence having at least 80% sequence identity thereto, more in particular a sequence of any one of SEQ ID NOs 1, 3 or 5 or a sequence having at least 80% sequence identity thereto; a sequence of any one of SEQ ID NOs 6, 8, 10, 12, 14, 15, 17, 18, 19, 20 or 22; a sequence having at least 60% sequence identity thereto; or any combination thereof.

[0101] In specific embodiments, Ntls and / or Ctls comprise at least 3, 4, 5, 6, 7, 8, 9 or 10 amino acid residues in total. Ntls and / or Ctls may, for example, comprise up to 30, 28, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11 or 10 amino acid residues in total.

[0102] In a specific embodiment, the stretch of amino acids present in comprises the sequence of any one of SEQ ID NOs 26 to 38 or a sequence having at least 80%, 85%, 90%, 95% or at least 98% sequence identity to any one of SEQ ID NOs 26 to 38

[0103] i) in case said heterologous amino acid sequence is incorporated into CDRH1, between amino acid residue HCres25 according to Kabat and the first amino acid residue of the natriuretic peptide;

[0104] ii) in case said heterologous amino acid sequence is incorporated into CDRH2, between amino acid residue HCres51 according to Kabat and the first amino acid residue of the natriuretic peptide;

[0105] iii) between amino acid residue HCres92 according to Kabat and the first amino acid residue of the natriuretic peptide in case said heterologous amino acid sequence is incorporated into CDRH3;

[0106] iv) in case said heterologous amino acid sequence is incorporated into CDRL1, between amino acid residue LCres26 according to Kabat and the first amino acid residue of the natriuretic peptide;

[0107] v) between amino acid residue LCres49 according to Kabat and the first amino acid residue of the natriuretic peptide in case said heterologous amino acid sequence is incorporated into CDRL2; and / or

[0108] vi) in case said heterologous amino acid sequence is incorporated into CDRL3, between amino acid residue LCres88 according to Kabat and the first amino acid residue of the natriuretic peptide.

[0109] In a specific embodiment, the stretch of amino acids between the last amino acid residue of the natriuretic peptide and the sequence of any one of SEQ ID NOs 39 to 51 or a sequence having at least 80%, 85%, 90%, 95% or at least 98% sequence identity to any one of SEQ ID NOs 39 to 51 is present.

[0110] i) amino acid residues HCres35a according to Kabat, in case said heterologous amino acid sequence is incorporated into CDRH1;

[0111] ii) amino acid residue HCres57 according to Kabat, in case said heterologous amino acid sequence is incorporated into CDRH2;

[0112] iii) amino acid residue HCres106 according to Kabat, in case said heterologous amino acid sequence is incorporated into CDRH3;

[0113] iv) amino acid residue LCres 32 according to Kabat in case said heterologous amino acid sequence is incorporated into CDRL1;

[0114] v) amino acid residue LCres57 according to Kabat, in case said heterologous amino acid sequence is incorporated into CDRL2; and / or

[0115] vi) amino acid residue LCres98 according to Kabat, in case said heterologous amino acid sequence is incorporated into CDRL3.

[0116] In a specific embodiment, the heterologous amino acid sequence consists of Ntls, natriuretic peptides, and Ctls.

[0117] In a specific embodiment, the stretch of amino acids present in comprises the sequence of any one of SEQ ID NOs 52 to 64 or a sequence having at least 80%, 85%, 90%, 95% or at least 98% sequence identity to any one of SEQ ID NOs 52 to 64

[0118] i) in case said heterologous amino acid sequence is incorporated into CDRH1, between amino acid residue HCres25 according to Kabat and amino acid residue HCres35a according to Kabat;

[0119] ii) in case said heterologous amino acid sequence is incorporated into CDRH2, between amino acid residue HCres51 according to Kabat and amino acid residue HCres57 according to Kabat;

[0120] iii) in case said heterologous amino acid sequence is incorporated into CDRH3, between amino acid residue HCres92 according to Kabat and amino acid residue HCres106 according to Kabat;

[0121] iv) in case said heterologous amino acid sequence is incorporated in CDRL1, between amino acid residue LCres26 according to Kabat and amino acid residue LCres 32 according to Kabat;

[0122] v) between amino acid residue LCres49 according to Kabat and amino acid residue LCres57 according to Kabat, in case said heterologous amino acid sequence is incorporated in CDRL2; and / or

[0123] vi) in case said heterologous amino acid sequence is incorporated in CDRL3, between amino acid residue LCres88 according to Kabat and amino acid residue LCres98 according to Kabat.

[0124] In a specific embodiment, the antibody or its fragment comprises at least two natriuretic peptides. In a specific embodiment, both natriuretic peptides are contained in a heterologous amino acid sequence that also comprises Ntls and Ctls and are incorporated into the CDR region of the antibody or its fragment as described herein. The at least two natriuretic peptides can be incorporated into two corresponding CDR regions of two light chains or two heavy chains, or the at least two natriuretic peptides can be incorporated into two separate CDR regions. The at least two natriuretic peptides can be the same or different. In a specific embodiment of this type, the antibody or its fragment comprises at least two different natriuretic peptides, which are incorporated into at least two CDR regions.

[0125] Due to the dimeric structure of the antibody molecule, a nucleic acid sequence encoding a heterologous amino acid sequence (Ntls-natriuretic peptide-Ctls) is inserted into the nucleic acid encoding the light chain or heavy chain of the immunoglobulin molecule, which will generally produce an antibody protein carrying two natriuretic peptides in the corresponding CDR regions of two identical light chains or two identical heavy chains. However, it is also contemplated that two nucleic acids encoding natriuretic peptides are inserted into two different CDR coding regions of the nucleic acid sequence encoding light and / or heavy chain, thereby producing an antibody molecule having four natriuretic peptides in two corresponding CDR pairs of the dimeric antibody. Also included are dimeric immunoglobulin molecules whose light and / or heavy chains are different, such as dimeric antibodies carrying a single natriuretic peptide and dimeric antibodies carrying two different natriuretic peptides in two corresponding CDR regions of two light chains or two heavy chains.

[0126] In specific embodiments, the natriuretic peptide is inserted into CDRH1 and CDRH2, CDRH1 and CDRH3, CDRH2 and CDRH3, CDRH1 and CDRL1, CDRH1 and CDRL2, CDRH1 and CDRL3, CDRH2 and CDRL1, or CDRH2 and CDRL2. In specific embodiments, the antibody or fragment thereof comprises one ANP and one BNP molecule; one ANP and one CNP molecule; or one BNP and one CNP molecule.

[0127] In a specific embodiment, the natriuretic peptide contained in at least one heterologous amino acid sequence incorporated into at least one CDR region of the antibody or its fragment is CNP, and the antibody or its fragment contains at least one other natriuretic peptide. In a specific embodiment, the at least one other natriuretic peptide is also contained in a heterologous amino acid sequence, which further contains Ntls and Ctls and is incorporated into the CDR region of the antibody or its fragment. In a specific embodiment, CNP and at least one other natriuretic peptide are incorporated into two corresponding CDR regions of two light chains or two heavy chains of the antibody or its fragment. In other specific embodiments, CNP and at least one other natriuretic peptide are incorporated into at least two separate CDR regions. Specifically, the at least one other natriuretic peptide is selected from ANP, BNP and CNP, more specifically selected from ANP and BNP.

[0128] An "empty" antibody molecule consisting of two heavy chains having the sequence of SEQ ID NO 65 and two light chains having the sequence of SEQ ID NO 66, which does not contain a heterologous amino acid sequence comprising a natriuretic peptide, is referred to as TPP-5657. In a specific embodiment, an antibody molecule consisting of two heavy chains having the sequence of SEQ ID NO 65, or a sequence having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, and two light chains having the sequence of SEQ ID NO 66, or a sequence having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, is used as the initial or parent antibody, into which a heterologous amino acid sequence comprising a natriuretic peptide is incorporated. In specific such embodiments, the heterologous amino acid sequence comprising a natriuretic peptide is incorporated into the CDR region of one or both heavy chains of such an antibody molecule.

[0129] In specific such embodiments, the heavy chain comprising at least one heterologous amino acid sequence incorporated into at least one of its CDR regions has the sequence of any one of SEQ ID NOs 67 to 79, or a sequence having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity to any one of SEQ ID NOs 67 to 79. In specific embodiments, an antibody of the invention is composed of two identical heavy chains having the sequence of any one of SEQ ID NOs 67 to 79, or a sequence having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity to any one of SEQ ID NOs 67 to 79, and two identical light chains having the sequence of SEQ ID NO 66, or a sequence having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto.

[0130] In specific other embodiments, a heterologous amino acid sequence comprising a natriuretic peptide is incorporated into the CDR region of one or both light chains of such antibody molecules. In specific such embodiments, the light chain comprising at least one heterologous amino acid sequence incorporated into at least one of its CDR regions has the sequence of any one of SEQ ID NOs 80 or 81, or a sequence having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity to any one of SEQ ID NOs 80 or 81. In specific embodiments, the antibodies of the invention are composed of two identical light chains having the sequence of any one of SEQ ID NOs 80 or 81, or a sequence having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity to any one of SEQ ID NOs 80 or 81, and two identical heavy chains having the sequence of SEQ ID NO 65, or a sequence having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto.

[0131] The following table describes the heavy and light chain composition of exemplary antibodies of the invention.

[0132] Table 1: Exemplary natriuretic peptide-grafted antibody constructs

[0133]

[0134]

[0135] The antibodies or fragments thereof of the present invention include naturally occurring purified products, products of chemical synthesis steps and products produced by recombinant technology. Based on their source, the antibodies or fragments thereof of the present invention may be glycosylated or non-glycosylated.

[0136] For example, standard recombinant DNA methods can be used to prepare and / or obtain nucleic acids encoding the heavy and light chains, incorporate these nucleic acids into expression vectors, and introduce the vectors into host cells for recombinant expression (see, e.g., Sambrook, Fritsch and Maniatis (eds.), Molecular Cloning; A Laboratory Manual, Second Edition, Cold Spring Harbor, NY, (1989); Ausubel, FM et al. (eds.) Current Protocols in Molecular Biology, Greene Publishing Associates, (1989); Goeddel, Gene Expression Technology, Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990); and U.S. Pat. No. 4,816,397 to Boss et al.).

[0137] Therefore, in a second aspect, the present invention relates to nucleic acids or nucleic acid mixtures encoding antibodies or fragments thereof of the present invention. These nucleic acid sequences may, in some cases, be optimized for mammalian expression. The DNA molecules of the present invention are not limited to the sequences disclosed herein, but also include variants thereof.

[0138] The present invention also provides a recombinant nucleic acid construct comprising one or more nucleic acid sequences of the present invention. The recombinant nucleic acid construct of the present invention may, for example, comprise a nucleic acid vector, such as a plasmid, into which a nucleic acid molecule encoding an antibody of the present invention or a fragment thereof has been inserted. It should be understood that the design of the expression vector (including the selection of regulatory sequences) is influenced by factors such as the choice of host cells, the desired protein expression level, and whether constitutive expression or inducible expression is desired.

[0139] The available expression vector used for bacterial use can be made up by inserting one or more nucleotide sequences of the present invention, suitable translation initiation and termination signal together with functional promoter into operable reading phase.Bacterial expression vector generally includes one or more phenotypic selection markers and replication origin, to ensure the maintenance of carrier and (if necessary) amplification is provided in bacterial host.Bacterial expression vector can comprise the element that is derived from commercially available plasmid, such as known cloning vector pBR322 (ATCC37017).According to the intended use of expressed antibody or its fragment, many bacterial expression vectors can be advantageously selected.For example, if a large amount of this class antibody is needed, it may be necessary to mediate the carrier of the high-level expression of the antibody fusion protein that is easy to purify.

[0140] The recombinant nucleic acid construct intended for antibody expression in a eukaryotic host cell may comprise regulatory sequences capable of regulating the expression of the open reading frame in the eukaryotic cell, preferably a promoter and a polyadenylation signal. Promoters and polyadenylation signals that are functional in the specific cell type intended for antibody expression are preferably selected. Examples of suitable promoters include, but are not limited to, promoters from cytomegalovirus (CMV) (e.g., the strong CMV immediate early promoter), simian virus 40 (SV40), mouse mammary tumor virus (MMTV), human immunodeficiency virus (HIV) (e.g., the HIV long terminal repeat (LTR) promoter), Moloney virus, Epstein-Barr virus (EBV), adenovirus (e.g., the adenovirus major late promoter (ADMLP)), polyoma and Rous sarcoma virus (RSV), a synthetic CAG promoter consisting of the CMV early enhancer element, the promoter, first exon and first intron of the chicken β-actin gene, and the splice acceptor of the rabbit β-globin gene; and promoters from mammalian genes such as actin, myosin, hemoglobin, muscle creatine, and metallothionein. In specific embodiments, the eukaryotic expression cassette comprises a CMV promoter. In the context of the present invention, the term "CMV promoter" refers to the strong immediate early cytomegalovirus promoter.

[0141] Examples of suitable polyadenylation signals include, but are not limited to, the bovine growth hormone (BGH) polyadenylation site, the SV40 polyadenylation signal, and the LTR polyadenylation signal.

[0142] In addition, the recombinant nucleic acid sequence may comprise one or more enhancer sequences. The enhancer may be, for example, an enhancer of mammalian actin, myosin, hemoglobin, muscle creatine, or a viral enhancer, such as an enhancer from CMV, RSV, SV40, or EBV. For further description of viral regulatory elements and sequences thereof, see US 5,168,062 of Stinski, US 4,510,245 of Bell et al., and US 4,968,615 of Schaffner et al.

[0143] Regulatory sequences and codons are generally species-dependent, so in order to maximize protein production, regulatory sequences and codons that are effective in the species / cell type intended for antibody expression are preferably selected. Those skilled in the art can produce recombinant DNA molecules that are functional in a given test species.

[0144] Mammalian recombinant expression vectors may also contain an origin of replication and a selectable marker (see, for example, US 4,399,216, US 4,634,665, and US 5,179,017). Suitable selectable markers include genes that confer resistance to drugs such as G418, puromycin, hygromycin, blasticidin, zeocin / bleomycin, or methotrexate, or selectable markers that utilize auxotrophic factors (such as glutamine synthetase) on the host cell into which the vector has been introduced. For example, the dihydrofolate reductase (DHFR) gene confers resistance to methotrexate, a novel gene confers resistance to G418, the bsd gene from Aspergillus terreus confers resistance to blasticidin, puromycin N-acetyltransferase confers resistance to puromycin, the Sh ble gene product confers resistance to bleomycin, and the Escherichia coli hygromycin resistance gene (hyg or hph) confers resistance to hygromycin. Selectable markers (such as DHFR or glutamine synthetase) can also be combined with MTX and MSX for amplification techniques.

[0145] In some embodiments, the nucleic acid sequence encoding heavy chain and light chain is inserted into a separate vector. In other embodiments, the nucleic acid sequence encoding heavy chain and light chain is inserted into the same vector. In addition, the nucleic acid sequence encoding the variable region of heavy chain and / or light chain can be converted to a nucleic acid sequence encoding, for example, a full-length antibody chain, a Fab fragment or scFv. The DNA fragment encoding VL or VH can be operably connected (so that the amino acid sequence encoded by the two DNA fragments is in frame) to another DNA fragment encoding, for example, an antibody constant region or a flexible linker. As an example, to create a polynucleotide sequence encoding an scFv, nucleic acids encoding VH and VL can be operably linked to another segment encoding a flexible linker so that the VH and VL sequences can be expressed as a continuous single-chain protein with the VL and VH regions connected by a flexible linker (see, e.g., Bird et al. (1988) Science 242:423-426; Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883; McCafferty et al., Nature (1990) 348:552-554). The sequences of human heavy and light chain constant regions are known in the art (see, e.g., 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), and DNA fragments encompassing these regions can be obtained by standard PCR amplification.

[0146] In a specific embodiment, the nucleic acid sequence encoding the heavy chain into which the heterologous amino acid sequence comprising a natriuretic peptide is incorporated comprises the sequence of any one of SEQ ID NOs 82 or 83, or a sequence having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity to any one of SEQ ID NOs 82 or 83. In specific such embodiments, the nucleic acid sequence encoding the light chain comprises the sequence of SEQ ID NO 84, or a sequence having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto.

[0147] In a third aspect, the present invention relates to host cells comprising nucleic acid of the present invention or nucleic acid mixtures. In the context of the present invention, the terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acids have been introduced, including the progeny of these cells. Host cells include "transformants," "converted cells," "transfectants," "transfected cells," and "transduced cells," which include cells of primary conversion / transfection / transduction and progeny derived therefrom, without considering passage number. The nucleic acid content of progeny may not be identical to that of the parental cell, and the exogenous nucleic acids included may contain mutations. This paper includes progeny with functions or biological activities identical to those screened or selected in the original transformed cell.

[0148] Transfection of the expression vector into the host cell can be performed using standard techniques, such as electroporation, nucleofection, calcium phosphate precipitation, lipofection, polycation-based transfection such as polyethyleneimine (PEI)-based transfection, and DEAE-dextran transfection.

[0149] Suitable host cells include prokaryotic cells and eukaryotic cells. Examples of prokaryotic host cells are, for example, bacteria, including but not limited to Escherichia coli, Bacillus subtilis, Salmonella typhimurium, and Pseudomonas, Streptomyces, and Staphylococcus.

[0150] Non-limiting examples of eukaryotic host cells include yeast, insects and insect cells, plants and plant cells, transgenic animals, and mammalian cells. Suitable mammalian host cells for antibody expression include Chinese hamster ovary (CHO cells), such as CHO-K1, CHO-S, CHO-K1SV (including dhfr-CHO cells described in Urlaub and Chasin, (1980) Proc. Natl. Acad. Sci. USA 77: 4216-4220 and Urlaub et al., Cell. 1983 Jun; 33 (2): 405-12, used with a DHFR selective marker, such as described in RJ Kaufman and PA Sharp (1982) Mol. Biol. 159: 601-621; and as described in Fan et al., Biotechnol Bioeng. 2012 Apr; 109(4): 1007-15), NS0 myeloma cells, COS cells, HEK293 cells, HKB11 cells, BHK21 cells, CAP cells, EB66 cells and SP2 cells.

[0151] Expression can also be transient or semi-stable in expression systems such as HEK293, HEK293T, HEK293-EBNA, HEK293E, HEK293-6E, HEK293-Freestyle, HKB11, Expi293F, 293EBNALT75, CHOFreestyle, CHO-S, CHO-K1, CHO-K1SV, CHOEBNALT85, CHOS-XE, CHO-3E7 or CAP-T cells (e.g., Durocher et al., Nucleic Acids Res. 2002 Jan 15;30(2):E9).

[0152] In a fourth aspect, the present invention relates to a method for producing an antibody or a fragment thereof, comprising culturing the host cell of the present invention. Specifically, the host cell of the present invention is cultured under conditions suitable for expression of the antibody or a fragment thereof.

[0153] Antibody expression can be constitutive or regulated (e.g., inducible). For inducible antibody expression, host cells of the present invention are typically grown to an appropriate cell density, and then the selected promoter is de-repressed / induced by an appropriate method (e.g., temperature shift or chemical induction, such as addition or removal of small molecule inducers, such as tetracycline that binds to the Tet system), and the host cells are cultured for an additional period of time.

[0154] In a specific embodiment, the method for producing an antibody of the present invention or its fragment also includes a step of recovering the antibody or its fragment from the host cell culture. The cells can be collected, for example, by centrifugation, and destroyed by physical or chemical methods, and the antibody or its fragment can be further purified from the resulting crude extract. In some embodiments, the expression vector is designed so that the expressed antibody or its fragment is secreted into the culture medium in which the host cells are grown. In this case, the antibody or its fragment can be directly recovered from the culture medium using standard protein purification methods.

[0155] In a specific embodiment, the method of the present invention further comprises a step of purifying and recovering the antibody or its fragment. Specifically, the antibody is purified (1) to greater than 90%, as determined, for example, by analytical chromatography or by SDS-capillary gel electrophoresis (e.g., on a Caliper LabChip GXII, GX90, or Biorad bioanalyzer), more specifically to produce an antibody homogeneity of at least about 92.5%, 95%, 98%, or 99%; or, the antibody is purified (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence, or (3) by SDS-PAGE to homogeneity under reducing or non-reducing conditions using Coomassie blue or, preferably, silver staining.

[0156] The antibodies or fragments thereof of the present invention can be recovered and purified from recombinant cell cultures by known methods, including but not limited to ammonium sulfate or ethanol precipitation, acid extraction, protein A chromatography, protein G chromatography, molecular sieve chromatography, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxyapatite chromatography, and lectin chromatography. High performance liquid chromatography ("HPLC") can also be used for purification (see, e.g., Colligan, Current Protocols in Immunology or Current Protocols in Protein Science, John Wiley & Sons, NY, NY, (1997-2001), e.g., Chapters 1, 4, 6, 8, 9, 10).

[0157] In a fifth aspect, the present invention relates to a composition comprising an antibody or fragment thereof of the present invention. Specifically, the composition of the present invention is a pharmaceutical composition suitable for use in a therapeutic method, wherein the antibody or fragment thereof of the present invention is contained in an amount effective to achieve the intended purpose, i.e., to prevent or treat a specific disease state.

[0158] The composition optionally further comprises at least one pharmaceutically acceptable excipient. In the context of the present invention, the term "excipient" refers to a natural or synthetic substance formulated with the active ingredient of the drug. Suitable excipients include antiadhesives, adhesives, coatings, disintegrants, spices, pigments, lubricants, glidants, sorbents, preservatives and sweeteners. Specific examples of pharmaceutically acceptable excipients include, but are not limited to, saline, buffered saline, dextrose and water. In the context of the present invention, the term "pharmaceutically acceptable" refers to other ingredients of a molecular entity and pharmaceutical composition that is physiologically tolerable and generally does not produce adverse reactions when administered to a mammal (e.g., a human). The term "pharmaceutically acceptable" may also refer to approval by a federal or state regulatory agency or listing in the U.S. Pharmacopoeia or other pharmacopoeias generally recognized for use in mammals (more specifically, humans).

[0159] The compositions of the present invention may also contain one or more other therapeutically active agents.

[0160] The pharmaceutical composition may be in the form of a solution, suspension, enteric-coated capsule, lyophilized powder, or any other form suitable for the intended use.

[0161] Pharmaceutical compositions for oral administration can be formulated using pharmaceutically acceptable excipients well known in the art in dosages suitable for oral administration. Such excipients enable the pharmaceutical compositions to be formulated into tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and the like for ingestion by a patient.

[0162] Pharmaceutical preparations for oral use can be obtained by combining the active compound with a solid excipient, optionally grinding the resulting mixture, and processing the granule mixture after adding suitable auxiliaries (if desired) to obtain tablets or dragee cores. Suitable excipients are carbohydrate or protein fillers, such as sugars, including lactose, sucrose, mannitol or sorbitol; starch from corn, wheat, rice, potato or other plants; cellulose, such as methylcellulose, hydroxypropyl methylcellulose or sodium carboxymethylcellulose; and gums, including gum arabic and tragacanth; and proteins, such as gelatin and collagen. If desired, disintegrants or solubilizers, such as cross-linked polyvinyl pyrrolidone, agar, alginic acid or a salt thereof, such as sodium alginate, may be added.

[0163] Dragee cores may be provided with suitable coatings such as concentrated sugar solutions, which may also contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol and / or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. Dyes or pigments may be added to the tablets or dragee coatings for product identification or to characterize the amount of active compound, i.e., the dosage.

[0164] Pharmaceutical preparations for oral administration include push-fit capsules made of gelatin and soft, sealed capsules made of gelatin and a coating such as glycerol or sorbitol. Push-fit capsules may contain the active ingredient mixed with a filler or binder such as lactose or starch, a lubricant such as talc or magnesium stearate, and optionally a stabilizer. In soft capsules, the active compound may be dissolved or suspended in a suitable liquid such as a fatty oil, liquid paraffin, or liquid polyethylene glycol, with or without a stabilizer.

[0165] Pharmaceutical preparations for parenteral administration include aqueous solutions of therapeutically active agents. For injection, pharmaceutical compositions of the present invention can be prepared in aqueous solutions, preferably in physiologically compatible buffers (such as Hank's solution, Ringer's solution or physiologically buffered saline). Aqueous injection suspensions can contain substances that increase suspension viscosity, such as sodium carboxymethyl cellulose, sorbitol or dextran. In addition, the suspension of active agent can be prepared as suitable oily injection suspensions. Suitable lipophilic solvents or carriers include fatty oils (such as sesame oil) or synthetic fatty acid esters (such as ethyl oleate or triglycerides) or liposomes. Optionally, the suspension can also contain suitable stabilizers or reagents that increase the solubility of the therapeutically active agent to allow the preparation of highly concentrated solutions.

[0166] For topical or nasal administration, penetrants appropriate to the particular barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

[0167] The pharmaceutical compositions of the present invention may be manufactured in a manner known in the art, eg, by means of conventional mixing, dissolving, granulating, dragee-making, suspending, emulsifying, encapsulating, entrapping or lyophilizing processes.

[0168] Pharmaceutical compositions can be provided in the form of salts and can be formed with acids including, but not limited to, hydrochloric acid, sulfuric acid, acetic acid, lactic acid, tartaric acid, malic acid, succinic acid, and the like. Salts tend to be more soluble in aqueous or other protic solvents than the corresponding free base forms. In other cases, a preferred preparation may be a lyophilized powder containing 1 mM-50 mM histidine, 0.1%-2% sucrose, 2%-7% mannitol, with a pH range of 4.5 to 7.5, which is combined with a buffer prior to use.

[0169] Further details of formulation and administration techniques may be found in the latest edition of Remington's Pharmaceutical Sciences (Ed. Maack Publishing Co, Easton, Pa.).

[0170] After preparing the pharmaceutical composition comprising the antibody or fragment thereof of the present invention, it can be placed in an appropriate container and labeled for the designated condition to be treated. For administration of the antibody or fragment of the present invention, such labeling will include the amount, frequency and method of administration.

[0171] The present invention also provides packages and kits comprising one or more containers containing one or more ingredients of the aforementioned compositions of the present invention, accompanied by a notification in a form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals or biological products reflecting that the agency has approved the manufacture, use, or sale of the product for human administration.

[0172] In a sixth aspect, the present invention relates to an antibody or fragment thereof of the present invention or a composition of the present invention for use in a method of treatment.

[0173] Specifically, this method of treatment involves administering a therapeutically effective amount of an antibody or fragment thereof of the present invention to a subject in need thereof. The subject can be a human or a non-human animal (such as a rabbit, rat, mouse, dog, monkey or other lower primate).

[0174] In the context of the present invention, the term "therapeutically effective amount" is defined as an amount of an antibody of the present invention or a fragment thereof, sufficient to prevent or alleviate the disease symptoms of any of the disorders and diseases mentioned herein, as a single dose or according to a multiple dose regimen, alone or in combination with other drugs. In specific embodiments, the "therapeutically effective amount" is toxicologically tolerable. The determination of an effective dose is well within the capabilities of those skilled in the art. The therapeutically effective amount of a therapeutic agent generally depends to a large extent on specific patient characteristics, such as age, weight, sex and disease state, the time, frequency and route of administration, the drug combination and the nature of the disorder being treated. Common dosages of antibodies range from 0.1 micrograms to 100,000 micrograms, up to a total dose of about 10 g, depending on the route of administration.

[0175] General guidance on how to determine this can be found, for example, in the publications of the International Conference on Harmonization and in Chapters 27 and 28 of REMINGTON'S PHARMACEUTICAL SCIENCES, pp. 484-528 (18th ed., Alfonso R. Gennaro, Ed., Easton, Pa.: Mack Pub. Co., 1990). More specifically, determining a therapeutically effective amount will depend on factors such as the toxicity and efficacy of the drug, which can be determined using methods well known in the art and described in the aforementioned references. In general, the therapeutic efficacy and toxicity of a therapeutic agent can be determined in cell culture assays or animal models, for example, as the ED50 (the dose therapeutically effective in 50% of a population) and the LD50 (the dose lethal to 50% of a population), respectively. The dose ratio of the ED50 to the LD50 is the therapeutic index.

[0176] The antibodies or fragments thereof of the present invention are useful for treating and / or preventing cardiovascular, renal, pulmonary, skeletal, ocular, thromboembolic and fibrotic diseases and disorders, dwarfism, achondroplasia, and other cGMP-related and / or natriuretic peptide-responsive disorders. Thus, in specific embodiments, the antibodies or fragments thereof are used to treat and / or prevent any one or any combination of these disorders and diseases.

[0177] Thus, the antibodies of the present invention or fragments thereof can be used in medicaments for the treatment and / or prevention of cardiovascular disorders, such as arterial and pulmonary hypertension, refractory and intractable hypertension, acute and chronic heart failure, coronary heart disease, bronchiolitis obliterans syndrome (BOS), tumor-related and oncological diseases, graft-versus-host disease, sickle cell disease, stable and unstable angina, peripheral and cardiovascular disorders, arrhythmias, atrial and ventricular arrhythmias and conduction impairment (such as atrioventricular block I-III degree (AB block I-III)). II)), supraventricular tachyarrhythmia, atrial fibrillation, atrial flutter, ventricular fibrillation, ventricular flutter, ventricular tachyarrhythmia, torsades de pointes, atrial and ventricular extrasystoles, AV junction extrasystoles, sick sinus syndrome, syncope, atrioventricular nodal reentrant tachycardia, Wolff-Parkinson-White syndrome, acute coronary syndrome (ACS), autoimmune heart disease (pericarditis, endocarditis, valvulitis, aortitis, cardiomyopathy), shock ( Such as cardiogenic shock, septic shock and anaphylactic shock), aneurysm, boxer cardiomyopathy (premature ventricular contraction (PVC)); can be used in drugs for the treatment and / or prevention of thromboembolic disorders and ischemia, such as myocardial ischemia, myocardial infarction, stroke, cardiac hypertrophy, transient ischemic attack, preeclampsia, inflammatory cardiovascular disorders, coronary artery and peripheral artery spasm, edema formation (such as pulmonary edema, cerebral edema, renal edema or edema caused by heart failure), peripheral circulatory disorders, reperfusion injury, arterial and venous thrombosis formation, microalbuminuria, myocardial insufficiency, endothelial dysfunction, prevention of restenosis after, for example, thrombolytic therapy, percutaneous transluminal angioplasty (PTA), transluminal coronary angioplasty (PTCA), heart transplantation and bypass surgery, and microvascular and macrovascular damage (vasculitis), increased fibrinogen and low-density lipoprotein (LDL) levels, increased plasminogen activator inhibitor 1 (PAI-1) concentrations; and in drugs for the treatment and / or prevention of erectile dysfunction and female sexual dysfunction.

[0178] In the context of the present invention, the term "heart failure" includes acute and chronic heart failure, and also includes more specific or related disease types such as acute decompensated heart failure, right heart failure, left heart failure, global heart failure, ischemic cardiomyopathy, dilated cardiomyopathy, hypertrophic cardiomyopathy, idiopathic cardiomyopathy, congenital heart defects, heart failure associated with heart valve defects, mitral stenosis, mitral insufficiency, aortic stenosis, aortic insufficiency, tricuspid stenosis, tricuspid insufficiency, pulmonary stenosis, pulmonary insufficiency, combined heart valve defects, myocardial inflammation (myocarditis), chronic myocarditis, acute myocarditis, viral myocarditis, diabetic heart failure, alcoholic cardiomyopathy, cardiac storage disorder, diastolic heart failure and systolic heart failure, heart failure with preserved ejection fraction (HFpEF), heart failure with reduced ejection fraction (HFrEF) and acute exacerbation of existing chronic heart failure (exacerbated heart failure).

[0179] In addition, the antibodies or fragments thereof of the present invention can also be used to treat and / or prevent arteriosclerosis, peripheral arterial disease (PAD), impaired lipid metabolism, hypolipoproteinemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia, hypercholesterolemia, abetalipoproteinemia, sitosterolemia, xanthomatosis, Tangier disease, adiposity, obesity, and combined hyperlipidemia and metabolic syndrome.

[0180] The antibodies or fragments thereof of the present invention can also be used to treat and / or prevent primary and secondary Raynaud's phenomenon, impaired microcirculation, claudication, peripheral and autonomic neuropathy, diabetic microangiopathy, diabetic retinopathy, diabetic limb ulcers, gangrene, CREST syndrome, erythromelalgia, onychomycosis, rheumatism, and to promote wound healing.

[0181] The antibodies or fragments thereof of the present invention are also suitable for the treatment of urinary disorders, such as benign prostatic syndrome (BPS), benign prostatic hyperplasia (BPH), benign prostatic hypertrophy (BPE), bladder outlet obstruction (BOO), lower urinary tract syndrome (LUTS, including feline urinary syndrome (FUS)), genitourinary disorders including neurogenic overactive bladder (OAB) and (IC), urinary incontinence (UI) (e.g., mixed urinary incontinence, urge urinary incontinence, stress urinary incontinence or overflow incontinence (MUI, UUI, SUI, OUI)), pelvic pain, benign and malignant disorders of the male and female genitourinary organs.

[0182] The antibodies or fragments thereof of the present invention are also suitable for treating and / or preventing renal disorders, in particular acute and chronic renal insufficiency and acute and chronic renal failure. In the context of the present invention, the term "renal insufficiency" includes acute and chronic manifestations of renal insufficiency, as well as potential or related renal disorders such as renal hypoperfusion, intradialytic hypotension, obstructive uropathy, glomerulopathy, glomerulonephritis, acute glomerulonephritis, glomerulosclerosis, tubulointerstitial disease, renal disorders (such as primary and congenital renal diseases), nephritis, immune renal disorders (such as renal transplant rejection and immune complex-induced renal disorders, toxic substance-induced nephropathy, contrast agent-induced nephropathy, diabetic and non-diabetic nephropathy, pyelonephritis, renal cysts, nephrosclerosis, hypertensive nephrosclerosis and nephrotic syndrome, thereof Diagnostic characteristics may include, for example, abnormally decreased creatinine and / or water excretion, abnormally elevated blood concentrations of urea, nitrogen, potassium, and / or creatinine, altered renal enzyme (e.g., glutamyl synthetase) activity, altered urine osmolality or urine volume, elevated microalbuminuria, macroalbuminuria, glomerular and arteriole lesions, renal tubular dilatation, hyperphosphatemia, and / or the need for dialysis. The present invention also encompasses the use of the antibodies or fragments thereof of the present invention for the treatment and / or prevention of sequelae of renal insufficiency, such as pulmonary edema, heart failure, uremia, anemia, electrolyte imbalances (e.g., hyperkalemia, hyponatremia), and disorders of bone and carbohydrate metabolism.

[0183] In addition, the antibodies or fragments thereof of the present invention are also suitable for treating and / or preventing asthmatic disorders, pulmonary arterial hypertension (PAH) and other forms of pulmonary hypertension (PH) including left heart disease, HIV, sickle cell anemia, thromboembolism (CTEPH), sarcoidosis, COPD or pulmonary fibrosis associated pulmonary hypertension, chronic obstructive pulmonary disease (COPD), acute respiratory distress syndrome (ARDS), acute lung injury (ALI), alpha-1-antitrypsin deficiency (AATD), pulmonary fibrosis, emphysema (e.g., smoking-induced emphysema), bronchiolitis obliterans syndrome (BOS) and cystic fibrosis (CF).

[0184] The antibodies or fragments thereof of the present invention are also suitable for controlling central nervous system disorders characterized by a disturbance in the NO / cGMP system. It is particularly suitable for improving perception, attention, learning or memory after cognitive impairment, such as those associated with, inter alia, mild cognitive impairment, age-related learning and memory impairment, age-related memory loss, vascular dementia, traumatic brain trauma, stroke, dementia occurring after a stroke (post-stroke dementia), post-traumatic brain trauma, general attention disorder, attention disorder in children with learning and memory problems, Alzheimer's disease, Lewy body dementia, frontal lobe degeneration dementia (including Pick's syndrome), Parkinson's disease, progressive nuclear palsy, corticobasal degeneration dementia, amyotrophic lateral sclerosis (ALS), Huntington's disease, demyelination, multiple sclerosis, thalamic degeneration, Creutzfeld-Jacob dementia, HIV dementia, schizophrenia or Korsakoff psychosis. It is also indicated for the treatment and / or prevention of central nervous system disorders such as anxiety, stress and depressive states, CNS-related sexual dysfunction and sleep disturbances, as well as for pathological disorders controlling the intake of food, stimulants and addictive substances.

[0185] In addition, the antibodies or fragments thereof of the present invention are also suitable for controlling cerebral blood flow and are therefore effective agents for controlling migraines. They are also suitable for preventing and controlling the sequelae of cerebral infarction (stroke), such as stroke, cerebral ischemia, and craniocerebral trauma. They can also be used to control pain and tinnitus.

[0186] In addition, the antibodies or fragments of the present invention have anti-inflammatory effects and can therefore be used as anti-inflammatory agents for the treatment and / or prevention of sepsis (SIRS), multiple organ failure (MODS, MOF), renal inflammatory disorders, chronic intestinal inflammation (IBD, Crohn's disease, UC), pancreatitis, peritonitis, rheumatoid diseases, inflammatory skin diseases and inflammatory eye diseases.

[0187] In addition, the antibodies or fragments thereof of the present invention can also be used to treat and / or prevent autoimmune diseases.

[0188] The antibodies or fragments thereof are also suitable for the treatment and / or prevention of fibrotic disorders of internal organs such as the lungs, heart, kidneys, reproductive system, bone marrow, in particular the liver, as well as cutaneous fibromas and fibrotic eye diseases. In the context of the present invention, the term fibrotic disorder particularly includes the following terms: liver fibrosis, cirrhosis, pulmonary fibrosis, endomyocardial fibrosis, nephropathy, glomerulonephritis, interstitial renal fibrosis, fibrotic lesions caused by diabetes, uterine fibromas, endometriosis, myelofibrosis and similar fibrotic disorders, scleroderma, morphea, keloids, hypertrophic scarring (also after surgical procedures), nevi, diabetic retinopathy, proliferative vitreoretinopathy and connective tissue disorders (e.g. sarcoidosis).

[0189] The antibodies or fragments thereof of the present invention are also useful in controlling postoperative scarring, such as that resulting from glaucoma surgery.

[0190] The antibodies or fragments thereof of the present invention can also be used cosmetically for aging and keratinized skin.

[0191] Furthermore, the antibodies or fragments thereof of the present invention are suitable for treating and / or preventing hepatitis, tumors, osteoporosis, glaucoma and gastroparesis.

[0192] In addition, the antibodies or fragments thereof of the present invention are suitable for treating and / or preventing ocular disorders, such as eye diseases responsive to natriuretic peptides, retinal disorders, glaucoma including primary open-angle glaucoma (POAG), angle-closure glaucoma, congenital / developmental glaucoma, retinopathy, ocular trauma, optic neuropathy, ocular hypertension, increased intraocular pressure, diabetic retinopathy, macular degeneration (AMD), age-related eye diseases, macular edema, scleritis, uveitis, dry eye, corneal epithelial abrasion, corneal ulcer.

[0193] In addition, the antibodies or fragments thereof of the present invention are suitable for treating bone and cartilage disorders, such as bone and cartilage diseases responsive to natriuretic peptides, arthritis, cartilage tissue degenerative diseases, osteoarthritis, cartilage degeneration, bone fractures, skeletal dysplasia, achondroplasia, osteoporosis, osteogenesis imperfecta, Paget's disease of bone (PDB), metabolic bone disease, age-related bone disease, osteomyelitis, osteonecrosis, rickets, osteomalacia, growth plate injuries and diseases, joint and bone replacement related defects, Marfan syndrome, sports injuries, muscular dystrophy, Duchenne muscular dystrophy.

[0194] Therefore, in another aspect, the present invention relates to the use of the antibodies of the present invention or fragments thereof for the treatment and / or prevention of disorders, in particular the disorders mentioned above.

[0195] In specific embodiments, the antibodies of the invention or fragments thereof can be used in methods for treating and / or preventing heart failure, angina pectoris, hypertension, pulmonary hypertension, ischemia, vascular disorders, renal insufficiency, thromboembolic disorders, fibrotic disorders, skeletal and bone disorders, ocular disorders, and arteriosclerosis.

[0196] In another aspect, the present invention relates to the use of an antibody or fragment thereof according to the present invention for the production of a medicament for the treatment and / or prevention of disorders, in particular the disorders mentioned above.

[0197] In a specific embodiment, the present invention relates to the use of an antibody or fragment thereof of the present invention for the production of a medicament for the treatment and / or prevention of heart failure, angina pectoris, hypertension, pulmonary hypertension, ischemia, vascular disorders, renal insufficiency, thromboembolic disorders, fibrotic disorders, dementia, arteriosclerosis, skeleton and bone disorders, eye disorders, dwarfism, achondroplasia and erectile dysfunction.

[0198] In another aspect, the present invention relates to a method for the treatment and / or prevention of disorders, in particular the disorders mentioned above, using an effective amount of at least one antibody or fragment thereof according to the present invention.

[0199] In a specific embodiment, the present invention relates to a method for treating and / or preventing heart failure, angina pectoris, hypertension, pulmonary hypertension, ischemia, vascular disorders, renal insufficiency, thromboembolic disorders, fibrotic disorders, tumors and oncological diseases, skeleton and bone disorders, eye disorders, dwarfism, achondroplasia and arteriosclerosis using an effective amount of at least one antibody of the present invention or fragment thereof.

[0200] The antibodies or fragments thereof of the present invention may be administered as the sole agent or in combination with one or more additional therapeutic agents. In some cases, the antibodies themselves may be modified. For example, the antibodies or fragments thereof may be conjugated to chemical entities, for example to further improve efficacy, stability and / or half-life. Specifically, the antibodies or fragments thereof of the present invention may be pegylated and / or hydroxyethyl starched (HESylated).

[0201] Thus, in a specific embodiment, the antibodies or fragments thereof of the present invention are used in a method of treatment in combination with at least one additional therapeutic agent, in particular for the purposes described above.

[0202] The present invention also provides a pharmaceutical combination comprising at least one antibody or fragment thereof of the present invention and at least one additional therapeutic agent.

[0203] In the context of the present invention, the term "pharmaceutical combination" is used in a manner known to those skilled in the art, and this combination may be a fixed combination, a non-fixed combination or a kit of parts.

[0204] In the context of the present invention, the term "fixed combination" is used in a manner known to the person skilled in the art and is defined as a combination wherein, for example, a first active ingredient (such as one or more antibodies or fragments thereof of the invention) and a further active ingredient are present together as one unit dose or as a single entity, such as a single dose formulation. An example of a "fixed combination" is a pharmaceutical composition wherein the first active ingredient and the further active ingredient are present as a mixture for simultaneous administration, such as in a formulation. Another example of a "fixed combination" is a pharmaceutical combination wherein the first active ingredient and the further active ingredient are present as one unit rather than as a mixture. Therefore, the present invention provides pharmaceutical compositions comprising at least one antibody or fragment thereof and at least one additional therapeutic agent, in particular for the treatment and / or prevention of the above-mentioned disorders.

[0205] In the context of the present invention, the terms "non-fixed combination" and "kit-of-parts" are used in a manner known to the person skilled in the art and are defined as a combination wherein the first active ingredient and the further active ingredient are present in more than one unit, for example in separate dosage preparations. An example of a non-fixed combination or kit-of-parts is a combination wherein the first active ingredient and the further active ingredient are present separately.

[0206] The components of the non-fixed combination or kit-of-parts may be administered separately, sequentially, simultaneously, concurrently or chronologically staggered.

[0207] The antibody of the present invention or its fragment can be administered simultaneously with, before or after the other therapeutically active agent. In the context of the present invention, the term "simultaneously" means that the antibody of the present invention or its fragment and at least one other therapeutically active agent are administered on the same day, more particularly within 12 hours, more particularly within 2 hours.

[0208] In a specific embodiment, the administration of the antibody or fragment thereof of the present invention and at least one other therapeutic agent occurs within eight consecutive weeks, more particularly within one to six consecutive weeks. The antibody or fragment thereof of the present invention and at least one other therapeutic agent can be administered by the same route or by different routes.

[0209] The antibodies or fragments thereof of the present invention can be combined, for example, with known agents from the same indication therapeutic group, such as agents for the treatment and / or prevention of diseases and conditions associated with hypertension, heart failure, pulmonary hypertension, COPD, asthma, cystic fibrosis, achondroplasia, hyperphosphatemia, chronic kidney disease (CKD), soft tissue calcification, chronic kidney disease-associated calcification, non-chronic kidney disease-associated calcification, medial calcification including Moenckeberg's medial sclerosis, and ulcerative colitis. sclerosis), atherosclerosis, intimal calcification, CKD-associated cardiac hypertrophy, CKD-associated renal dystrophy, osteoporosis, postmenopausal osteoporosis, type 2 diabetes mellitus, chronic kidney disease, aging, hypophosphaturia, hyperparathyroidism, vitamin D disorders, vitamin K deficiency, vitamin K antagonist coagulants, Kawasaki disease, ACDC (arterial calcification due to CD73 deficiency), GACI (generalized arterial calcification of infancy), IBGC (idiopathic basal ganglia calcification), PXE (pseudoxanthoma elasticum), rheumatoid arthritis, Singleton-Merten syndrome, P-thalassemia, calciphylaxis, heterotrophic ossification, placental calcification of prematurity, uterine calcification, calcified uterine fibroids, Morbus Fahr disease, aortic valve microcalcifications, and calcifications.

[0210] Preferred examples of suitable other therapeutic agents to be combined with the antibodies or fragments thereof of the present invention:

[0211] - Organic nitrates and NO donors, such as sodium nitroprusside, nitroglycerin, isosorbide mononitrate, isosorbide dinitrate, molsidomide, or SIN-1, and inhaled NO;

[0212] Compounds that inhibit the breakdown of cyclic guanosine monophosphate (cGMP), for example inhibitors of phosphodiesterase (PDE) 1, 2 and / or 5, in particular PDE 5 inhibitors, such as sildenafil, vardenafil, tadalafil, udenafil, desantafil, avanafil, mirodenafil, lodenafil or PF-00489791;

[0213] - antithrombotic agents, for example and preferably platelet aggregation inhibitors, anticoagulants or profibrinolytic substances;

[0214] - antihypertensive active ingredients, for example and preferably calcium antagonists, angiotensin AII antagonists, ACE inhibitors, NEP inhibitors, vasopeptidase inhibitors, endothelin antagonists, renin inhibitors, alpha-blockers, beta-blockers, mineralocorticoid receptor antagonists, rho-kinase inhibitors and diuretics;

[0215] antiarrhythmic agents, for example and preferably sodium channel blockers, beta-blockers, potassium channel blockers, calcium antagonists, If channel blockers, digitalis, vagoliytics, sympathomimetics and other antiarrhythmic drugs, such as adenosine, adenosine receptor agonists and vernakalant;

[0216] -Positive inotropic agents, such as cardiac glycosides (Dogoxin), beta-adrenergic and dopaminergic agonists, such as isoproterenol, epinephrine, norepinephrine, dopamine, or dobutamine;

[0217] - vasopressin receptor antagonists, for example and preferably conivaptan, tolvaptan, lixivaptan, mozavaptan, satavaptan, SR-121463, RWJ 676070 or BAY 86-8050 and the compounds described in WO 2010 / 105770, WO 2011 / 104322 and WO 2016 / 071212;

[0218] - active ingredients that alter lipid metabolism, for example and preferably thyroid receptor agonists, PCSK9 inhibitors, cholesterol synthesis inhibitors, for example and preferably HMG-CoA reductase inhibitors or squalene synthesis inhibitors, ACAT inhibitors, CETP inhibitors, MTP inhibitors, PPAR-alpha, PPAR-gamma and / or PPAR-delta agonists, cholesterol absorption inhibitors, lipase inhibitors, polymeric bile acid sorbents, bile acid reabsorption inhibitors and lipoprotein (a) antagonists.

[0219] anti-inflammatory agents, for example and preferably glucocorticoids, for example and preferably prednisone, prednisolone, methylprednisolone, triamcinolone, dexamethasone, beclomethasone, betamethasone, flunisolide, budesonide or fluticasone and nonsteroidal anti-inflammatory agents (NSAIDs), for example and preferably acetylsalicylic acid (aspirin), ibuprofen and naproxen, 5-aminosalicylic acid derivatives, leukotriene antagonists, TNF-α inhibitors and chemokine receptor antagonists, such as CCR1, 2 and / or 5 inhibitors;

[0220] - agents which inhibit signal transduction cascades, such as and preferably kinase inhibitors, such as and preferably tyrosine kinase and / or serine / threonine kinase inhibitors;

[0221] agents that inhibit the degradation and modification of the extracellular matrix, for example and preferably inhibitors of matrix metalloproteinases (MMPs), for example and preferably inhibitors of chymotrypsin, stromelysine, collagenase, gelatinase and aggrecanase (preferably MMP-1, MMP-3, MMP-8, MMP-9, MMP-10, MMP-11 and MMP-13), and inhibitors of metalloelastase (MMP-12) and neutrophil elastase (HNE), for example sivelestat or DX-890;

[0222] - agents that block the binding of serotonin to its receptors, for example and preferably 5-HT2b receptor antagonists;

[0223] - antifibrotic agents, for example and preferably nintedanib, pirfenidone, adenosine A2b receptor antagonists, sphingosine-1-phosphate receptor 3 (S1P3) antagonists, autotaxin inhibitors, lysophosphatidic acid receptor 1 (LPA-1) and lysophosphatidic acid receptor 2 (LPA-2) antagonists, lysyl oxidase (LOX) inhibitors, lysyl oxidase-like 2 inhibitors, CTGF inhibitors, IL-13 antagonists, integrin antagonists, TGF-β antagonists, wnt signaling inhibitors, CCR2-antagonists;

[0224] - agents acting as bronchodilators, for example and preferably 5-HT2b receptor antagonists; β2-adrenergic agonists (short- and long-acting), anticholinergics and theophylline;

[0225] - agents that are antagonists of cytokines and chemokines, for example and preferably antagonists of TGF-β, CTGF, IL-1, IL-4, IL-5, IL-6, IL-8, IL-13, IL-25, IL-33, TSLP and integrins;

[0226] - organic nitrates and NO donors, for example and preferably sodium nitroprusside, nitroglycerin, isosorbide mononitrate, isosorbide dinitrate, molsidomine or SIN-1, and inhaled NO;

[0227] NO-independent, heme-dependent stimulators of soluble guanylate cyclase, such as, for example and preferably, the compounds described in WO 00 / 06568, WO 00 / 06569, WO 02 / 42301, WO 03 / 095451, WO 2011 / 147809, WO 2012 / 004258, WO 2012 / 028647 and WO 2012 / 059549;

[0228] NO-independent and heme-independent activators of soluble guanylate cyclase, such as, for example and preferably, the compounds described in WO 01 / 19355, WO 01 / 19776, WO 01 / 19778, WO 01 / 19780, WO 02 / 070462 and WO 02 / 070510;

[0229] - agents which stimulate cGMP synthesis, such as sGC modulators, for example and preferably riociguat, cinaciguat, vericiguat;

[0230] a prostacyclin analogue or an IP receptor agonist, for example and preferably iloprost, beraprost, treprostinil, epoprostenol or Selexipag;

[0231] - an endothelin receptor antagonist, for example and preferably bosentan, darusentan, ambrisentan or sitaxsentan;

[0232] - agents which inhibit soluble epoxide hydrolases (sEH), for example and preferably N,N′-di-cyclohexylurea, 12-(3-adamantan-1-yl-ureido)-dodecanoic acid or 1-adamantan-1-yl-3-{5-[2-(2-ethoxyethoxy)ethoxy]pentyl}-urea;

[0233] - agents that interact with glucose metabolism, for example and preferably insulin, biguanides, thiazolidinediones, sulfonylureas, acarbose, DPP4 inhibitors, GLP-1 analogues or SGLT-1 inhibitors;

[0234] - a natriuretic peptide, for example and preferably atrial natriuretic peptide (ANP, caperitide), brain natriuretic peptide (BNP, nesiritide), C-type natriuretic peptide (CNP) or urodilatin;

[0235] - natriuretic peptide derivatives, for example and preferably vosoritide, cenderitide, PL3994;

[0236] - cardiac myosin activators, for example and preferably omecamtiv mecarbil (CK-1827452);

[0237] - a calcium sensitizer, for example and preferably levosimendan;

[0238] - agents that influence cardiac energy metabolism, for example and preferably etomoxir, dichloroacetate, ranolazine or trimetazidine, full or partial adenosine A1 receptor agonists, such as GS-9667 (formerly known as CVT-3619), capadenoson and neladenoson;

[0239] - Agents that influence heart rate, for example and preferably ivabradine.

[0240] Antithrombotic agents are preferably understood to be compounds from the group of platelet aggregation inhibitors, anticoagulants or profibrinolytic substances.

[0241] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a platelet aggregation inhibitor, such as, for example and preferably, aspirin, clopidogrel, prasugrel, ticagrelor, ticlopidine or dipyridamole.

[0242] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a thrombin inhibitor, such as for example and preferably ximelagatran, dabigatran, melagatran, bivalirudin or clexane.

[0243] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a GPIIb / IIIa antagonist, such as, for example and preferably, tirofiban or abciximab.

[0244] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a factor Xa inhibitor, for example and preferably rivaroxaban, DU-176b, apixaban, betrixaban, otamixaban, fidexaban, razaxaban, letaxaban, eribaxaban, fondaparinux, idraparinux, PMD-3112, darexaban (YM-150), KFA-1982, EMD-503982, MCM-17, MLN-1021, DX 9065a, DPC 906, JTV803, SSR-126512 or SSR-128428.

[0245] In a specific embodiment, an antibody of the invention or fragment thereof is administered in combination with heparin or a low molecular weight (LMW) heparin derivative.

[0246] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a vitamin K antagonist, such as, for example and preferably, coumarin.

[0247] Antihypertensive agents are preferably understood to be compounds from the group of calcium antagonists, angiotensin AII antagonists, ACE inhibitors, endothelin antagonists, renin inhibitors, alpha-receptor blockers, beta-receptor blockers, mineralocorticoid receptor antagonists, rho-kinase inhibitors and diuretics.

[0248] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a calcium antagonist, such as for example and preferably nifedipine, amlodipine, verapamil or diltiazem.

[0249] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an alpha-1 receptor blocker, such as, for example and preferably, prazosin.

[0250] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a beta-blocker, for example and preferably propranolol, atenolol, timolol, pindolol, alprenolol, oxprenolol, penbutolol, bupranolol, metipranolol, nadolol, mepindolol, carazolol, sosulfamethoxazole, metoprolol, betaxolol, celiprolol, bisoprolol, carteolol, esmolol, labetalol, carvedilol, adalolol, landiolol, nebivolol, epanolol or bucindolol.

[0251] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an angiotensin AII antagonist, for example and preferably losartan, candesartan, valsartan, telmisartan or embusartan, or a dual angiotensin AII antagonist / neprilysin inhibitor, for example and preferably LCZ696 (valsartan / sacubitril).

[0252] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an ACE inhibitor, such as for example and preferably enalapril, captopril, lisinopril, ramipril, delapril, fosinopril, quinapril, perindopril or quinapril.

[0253] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an endothelin antagonist, such as for example and preferably bosentan, darusentan, ambrisentan or sitaxsentan.

[0254] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a renin inhibitor, for example and preferably aliskiren, SPP-600 or SPP-800.

[0255] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a mineralocorticoid receptor antagonist, such as for example and preferably finerenone, spironolactone or eplerenone.

[0256] In specific embodiments, the antibodies or fragments thereof of the invention are administered in combination with loop diuretics, such as furosemide, torsemide, bumetanide, and piretanide, with potassium-sparing diuretics, such as amiloride and triamterene, with aldosterone antagonists, such as spironolactone, potassium canrenoate, and eplerenone, and with thiazide diuretics, such as hydrochlorothiazide, chlorthalidone, xipamide, and indapamide.

[0257] Lipid metabolism modifiers are preferably understood to be compounds from the group of CETP inhibitors, thyroid receptor agonists, cholesterol synthesis inhibitors such as HMG-CoA reductase inhibitors or squalene synthesis inhibitors, ACAT inhibitors, MTP inhibitors, PPAR-alpha, PPAR-gamma and / or PPAR-delta agonists, cholesterol absorption inhibitors, polymeric bile acid sorbents, bile acid reabsorption inhibitors, lipase inhibitors and lipoprotein (a) antagonists.

[0258] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a CETP inhibitor, for example and preferably dalcetrapib, anacetrapib, torcetrapib (CP-529414), JJT-705 or CETP vaccine (Avant).

[0259] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a thyroid receptor agonist, such as, for example and preferably, D-thyroxine, 3,5,3'-triiodothyronine (T3), CGS 23425 or acitreol (CGS26214).

[0260] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an HMG-CoA reductase inhibitor from the statin class, such as for example and preferably lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin, rosuvastatin or pitavastatin.

[0261] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a squalene synthesis inhibitor, such as, for example and preferably, BMS-188494 or TAK-475.

[0262] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an ACAT inhibitor, such as for example and preferably avasimibe, melinamide, pactimibe, eflucimibe or SMP-797.

[0263] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an MTP inhibitor, such as for example and preferably implitapid, BMS-201038, R-103757 or JTT-130.

[0264] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a PPAR-γ agonist, such as, for example and preferably, pioglitazone or rosiglitazone.

[0265] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a PPAR-delta agonist, such as for example and preferably GW 501516 or BAY 68-5042.

[0266] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a cholesterol absorption inhibitor, such as for example and preferably ezetimibe, tiqueside or pamaside.

[0267] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a lipase inhibitor, such as, for example and preferably, orlistat.

[0268] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a polymeric bile acid sorbent, such as, for example and preferably, cholestyramine, colestipol, colesolvam, cholesteat or colestimide.

[0269] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a bile acid reabsorption inhibitor, for example and preferably an ASBT (= IBAT) inhibitor, such as AZD-7806, S-8921, AK-105, BARI-1741, SC-435 or SC-635.

[0270] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a lipoprotein(a) antagonist, such as, for example and preferably, gemcabene calcium (CI-1027) or niacin.

[0271] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a lipoprotein(a) antagonist, such as, for example and preferably, gemcabene calcium (CI-1027) or niacin.

[0272] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a sGC modulator, for example and preferably riociguanidine, cinaciguanidine or veraciroxguanidine.

[0273] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an agent that influences glucose metabolism, such as, for example and preferably, insulin, sulfonylurea, acarbose, a DPP4 inhibitor, a GLP-1 analogue or an SGLT-1 inhibitor.

[0274] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a TGFβ antagonist, such as, for example and preferably, pirfenidone, nintedanib or fresolimumab.

[0275] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a CCR2 antagonist, for example and preferably CCX-140.

[0276] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a TGFα antagonist, such as, for example and preferably, adalimumab.

[0277] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a galectin-3 inhibitor, for example and preferably GCS-100.

[0278] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an Nrf-2 inhibitor, such as, for example and preferably, bardoxolone.

[0279] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a BMP-7 agonist, such as and preferably THR-184.

[0280] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a NOX1 / 4 inhibitor, such as, for example and preferably, GKT-137831.

[0281] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an agent that influences vitamin D metabolism, such as, for example and preferably, calcitriol, alfacalcidol, doxercalciferol, maxacalcitol, paricalcitol, cholecalciferol or poncatinol.

[0282] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a cytostatic agent, such as, for example and preferably, cyclophosphamide.

[0283] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an immunosuppressive agent, for example and preferably cyclosporine.

[0284] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a phosphate binder, such as, for example and preferably, colestilan, sevelamer hydrochloride and sevelamer carbonate, lanthanum and lanthanum carbonate.

[0285] In a specific embodiment, an antibody of the invention or fragment thereof is administered in combination with a renal proximal tubule sodium phosphate co-transporter, such as and preferably niacin or niacinamide.

[0286] In a specific embodiment, the antibodies or fragments thereof of the present invention are administered in combination with a calcimimetic for the treatment of hyperparathyroidism.

[0287] In a specific embodiment, the antibodies or fragments thereof of the invention are administered in combination with an agent for the treatment of iron deficiency, such as, and preferably, an iron product.

[0288] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an agent for the treatment of hyperuricaemia, such as, for example and preferably, allopurinol or rasburicase.

[0289] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a glycoprotein hormone for the treatment of anemia, such as, for example and preferably, erythropoietin.

[0290] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a biologic for immunotherapy, such as, for example and preferably, abatacept, rituximab, eculizumab or belimumab.

[0291] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a vasopressin antagonist for the treatment of heart failure (vaptanes), for example and preferably tolvaptan, conivaptan, lixivaptan, mozavaptan, satavaptan or relcovaptan.

[0292] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a Jak inhibitor, for example and preferably ruxolitinib, tofacitinib, baricitinib, CYT387, GSK2586184, lestaurtinib, pacritinib (SB1518) or TG101348.

[0293] In a specific embodiment, the antibodies of the present invention or fragments thereof are administered in combination with a prostacyclin analog for the treatment of microthrombi.

[0294] In a specific embodiment, an antibody of the invention or fragment thereof is administered in combination with a base treatment, for example and preferably sodium bicarbonate.

[0295] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an mTOR inhibitor, such as, for example and preferably, everolimus or rapamycin.

[0296] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an NHE3 inhibitor, for example and preferably AZD1722 or tenapanor.

[0297] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with an eNOS modulator, such as, and preferably, sapropterin.

[0298] In a specific embodiment, the antibodies of the invention or fragments thereof are administered in combination with a CTGF inhibitor, for example and preferably FG-3019.

[0299] One or more physiologically acceptable carriers or excipients can be used to formulate the antibody or its fragment for the treatment method of the present invention in any conventional manner. The antibody or its fragment of the present invention can be given by any appropriate means, which can be changed according to the type of obstacle to be treated. Possible routes of administration include enteral (e.g., oral), parenteral (e.g., intravenous, intraarterial, intraperitoneal, intramuscular, subcutaneous, intracardial, intraventricular, intrathecal, intramedullary, intralesional), intrapulmonary, and intranasal administration. In addition, the antibody of the present invention or its fragment can be administered by vein infusion, for example, with the dosage of the antibody or its fragment reduced. Preferably, by injection, most preferably by intravenous or subcutaneous administration, it depends in part on whether the disease is acute or chronic. The amount to be administered will depend on multiple factors, such as individual clinical symptoms, sex, age and / or body weight, whether to give other drugs, etc. The technician will recognize that the route of administration will change according to the obstacle to be treated or the disease.

[0300] Parenteral delivery methods include topical, intraarterial, intratumoral, intramuscular, subcutaneous, intramedullary, intrathecal, intracerebroventricular, intravenous, intraperitoneal, or intranasal administration.

[0301] In a specific embodiment, the method of treatment comprises single or multiple administrations of an antibody or fragment thereof or a pharmaceutical composition comprising the antibody or fragment thereof. The single doses may be the same or different. Specifically, the method of treatment comprises 1, 2, 3, 4, 5 or 6 administrations of an antibody or fragment thereof of the invention, preferably wherein the multiple administrations occur over a continuous period of one to six months. The antibody or fragment thereof of the invention may be administered, for example, every 3 to 4 days, weekly, once every two weeks or once every three weeks, depending on its half-life and clearance rate. Sequence Listing <110> Bayer AG <120> C-type natriuretic peptide grafted antibody <130> BHC 18 1 018 <160> 447 <170> PatentIn version 3.5 <210> 1 <211> 10 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 1 Ser Tyr Ala Met Ser Trp Val Arg Gln Ala 1 5 10 <210> 2 <211> 10 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 2 Thr Tyr Ile Ser Asn Val Asn His Lys Pro 1 5 10 <210> 3 <211> 9 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 3 Ser Asn Thr Lys Val Asp Lys Lys Val 1 5 <210> 4 <211> 10 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 4 Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly 1 5 10 <210> 5 <211> 9 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 5 Asp Arg Phe Ser Gly Ser Lys Ser Gly 1 5 <210> 6 <211> 15 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 6 Ser Ala Leu Leu Lys Ser Lys Leu Arg Ala Leu Leu Thr Ala Pro 1 5 10 15 <210> 7 <211> 14 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 7 Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp Gly 1 5 10 <210> 8 <211> 17 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 8 Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Asp Val Trp Gly 1 5 10 15 Gln <210> 9 <211> 12 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 9 Ser Val Val Val Thr Ser His Gln Ala Pro Gly Glu 1 5 10 <210> 10 <211> 10 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 10 Gly Glu Lys Lys Lys Leu Lys Ser Leu Ala 1 5 10 <210> 11 <211> 10 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 11 Gln Val Lys Leu Glu Leu Gly His Arg Ala 1 5 10 <210> 12 <211> 10 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 12 Val Asn His Leu Arg Ser Glu Lys Leu Thr 1 5 10 <210> 13 <211> 7 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 13 Ala Val Val Asn Val Arg Ala 1 5 <210> 14 <211> 7 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 14 Lys Gly Asp Lys Ile Ala Ile 1 5 <210> 15 <211> 10 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 15 Val Pro Lys Glu Lys Glu Lys Glu Lys Val 1 5 10 <210> 16 <211> 12 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 16 Ala Thr Lys Ala Val Ser Val Leu Lys Gly Asp Gly 1 5 10 <210> 17 <211> 9 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 17 Gln Gly Ile Ile Asn Phe Glu Gln Lys 1 5 <210> 18 <211> 11 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 18 Gly Pro Val Lys Val Trp Gly Ser Ile Lys Gly 1 5 10 <210> 19 <211> 7 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 19 Tyr Glu Lys Glu Lys Glu Lys 1 5 <210> 20 <211> 10 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> 20 Gly Gly Ser Thr Pro Leu Lys Ser Leu Ala 1 5 10 <210> twenty one <211> 14 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> twenty one Gly Ile Thr Gly Thr Lys Lys Tyr Gln Ser Ser Pro Asp Gly 1 5 10 <210> twenty two <211> 17 <212> PRT <213> Artificial <220> <223> Adapter sequence <400> twenty two Ser Tyr Ser Tyr Thr Tyr Asn Tyr Ala Glu Tyr Phe Gln His Trp Gly 1 5 10 15 Gln <210> twenty three <211> 28 <212> PRT <213> people <400> twenty three Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met Asp Arg Ile Gly 1 5 10 15 Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr 20 25 <210> twenty four <211> 28 <212> PRT <213> people <400> twenty four Met Val Gln Gly Ser Gly Cys Phe Gly Arg Lys Met Asp Arg Ile Ser 1 5 10 15 Ser Ser Ser Gly Leu Gly Cys Lys Val Leu Arg Arg 20 25 <210> 25 <211> twenty two <212> PRT <213> people <400> 25 Gly Leu Ser Lys Gly Cys Phe Gly Leu Lys Leu Asp Arg Ile Gly Ser 1 5 10 15 Met Ser Gly Leu Gly Cys 20 <210> 26 <211> 15 <212> PRT <213> Artificial <220> <223> N-terminal sequence (excluding reference aa) <400> 26 Gly Phe Thr Phe Gly Ser Gly Ser Gly Ser Gly Gly Ser Gly Gly 1 5 10 15 <210> 27 <211> twenty three <212> PRT <213> Artificial <220> <223> N-terminal sequence (excluding reference aa) <400> 27 Gly Phe Thr Phe Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 1 5 10 15 Pro Asp Gly Gly Ser Gly Gly 20 <210> 28 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (excluding reference aa) <400> 28 Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 29 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (excluding reference aa) <400> 29 Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 30 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (excluding reference aa) <400> 30 Ser Gly Pro Asn Pro Asn Lys Asn Pro Asn Pro Asn Pro Gly Ser Gly 1 5 10 15 Gly <210> 31 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (excluding reference aa) <400> 31 Ser Gly Pro Asn Pro Asn Pro Asn Pro Asn Pro Asn Ser Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 32 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (excluding reference aa) <400> 32 Ala Lys Val His Pro Asn Pro Asn Pro Asn Pro Asn Ser Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 33 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (excluding reference aa) <400> 33 Ala Ala Trp Asn Pro Asn Pro Asn Pro Asn Pro Asn Pro Asn Gly Gly 1 5 10 15 Ser Gly Gly <210> 34 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (excluding reference aa) <400> 34 Ala Lys Gly Ile Thr Gly Thr Lys Lys Tyr Gln Ser Ser Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 35 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (excluding reference aa) <400> 35 Ser Gly Ser Thr Tyr Ile Ser Asn Val Asn His Lys Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 36 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (excluding reference aa) <400> 36 Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Pro Asp Gly Gly Ser 1 5 10 15 Gly Gly <210> 37 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (excluding reference aa) <400> 37 Gly Val Pro Lys Glu Lys Glu Lys Glu Lys Val Ser Thr Ala Val Gly 1 5 10 15 Gly <210> 38 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (excluding reference aa) <400> 38 Ser Val Val Val Thr Ser His Gln Ala Pro Gly Glu Gly Gly Ser Gly 1 5 10 15 Gly <210> 39 <211> 11 <212> PRT <213> Artificial <220> <223> C-terminal sequence (excluding reference aa) <400> 39 Gly Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly 1 5 10 <210> 40 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (excluding reference aa) <400> 40 Gly Ser Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly 1 5 10 <210> 41 <211> 17 <212> PRT <213> Artificial <220> <223> C-terminal sequence (excluding reference aa) <400> 41 Gly Ser Tyr Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly Asn Arg Pro 1 5 10 15 Ser <210> 42 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (excluding reference aa) <400> 42 Gly Ser Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 1 5 10 15 <210> 43 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (excluding reference aa) <400> 43 Gly Ser Asn Pro Asn Glu Asn Pro Asn Pro Asn Pro Gly Ser 1 5 10 <210> 44 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (excluding reference aa) <400> 44 Gly Ser Tyr Asn Pro Asn Pro Asn Pro Asn Pro Asn Pro Gly Ser 1 5 10 15 <210> 45 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (excluding reference aa) <400> 45 Gly Ser Tyr Asn Pro Asn Pro Asn Pro Asn Pro His Val Asp Val Trp 1 5 10 15 Gly Gln <210> 46 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (excluding reference aa) <400> 46 Gly Ser Asn Pro Asn Pro Asn Pro Asn Pro Asn Pro Asn Val 1 5 10 <210> 47 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (excluding reference aa) <400> 47 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Ala Glu Tyr Phe Gln His Trp 1 5 10 15 Gly Gln <210> 48 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (excluding reference aa) <400> 48 Gly Ser Asn Thr Lys Val Asp Lys Lys Val Glu Gly Gly Ser 1 5 10 <210> 49 <211> 13 <212> PRT <213> Artificial <220> <223> C-terminal sequence (excluding reference aa) <400> 49 Gly Ser Asp Arg Phe Ser Gly Ser Lys Ser Gly Gly Ser 1 5 10 <210> 50 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (excluding reference aa) <400> 50 Gly Ser Ala Pro Leu Glu Val Pro Lys Glu Lys Glu Lys Glu Lys Val 1 5 10 15 <210> 51 <211> 11 <212> PRT <213> Artificial <220> <223> C-terminal sequence (excluding reference aa) <400> 51 Gly Gly Ser Thr Pro Leu Lys Ser Leu Ala Ser 1 5 10 <210> 52 <211> 54 <212> PRT <213> Artificial <220> <223> N-terminal sequence-NP-C-terminal sequence (excluding reference aas) <400> 52 Gly Phe Thr Phe Gly Ser Gly Ser Gly Ser Gly Gly Ser Gly Gly Ser 1 5 10 15 Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met Asp Arg Ile Gly Ala 20 25 30 Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr Gly Gly Ser Gly Gly 35 40 45 Ser Gly Ser Gly Ser Gly 50 <210> 53 <211> 65 <212> PRT <213> Artificial <220> <223> N-terminal sequence-NP-C-terminal sequence (excluding reference aas) <400> 53 Gly Phe Thr Phe Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 1 5 10 15 Pro Asp Gly Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly 20 25 30 Gly Arg Met Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser 35 40 45 Phe Arg Tyr Gly Ser Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 50 55 60 Gly 65 <210> 54 <211> 64 <212> PRT <213> Artificial <220> <223> N-terminal sequence-NP-C-terminal sequence (excluding reference aas) <400> 54 Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met Asp 20 25 30 Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr Gly 35 40 45 Ser Tyr Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly Asn Arg Pro Ser 50 55 60 <210> 55 <211> 63 <212> PRT <213> Artificial <220> <223> N-terminal sequence - NP - C-terminal sequence (excluding reference aas) <400> 55 Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met 20 25 30 Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr 35 40 45 Gly Ser Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 50 55 60 <210> 56 <211> 59 <212> PRT <213> Artificial <220> <223> N-terminal sequence-NP-C-terminal sequence (wo ref aas) <400> 56 Ser Gly Pro Asn Pro Asn Lys Asn Pro Asn Pro Asn Pro Gly Ser Gly 1 5 10 15 Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met Asp Arg Ile 20 25 30 Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr Gly Ser Asn 35 40 45 Pro Asn Glu Asn Pro Asn Pro Asn Pro Gly Ser 50 55 <210> 57 <211> 63 <212> PRT <213> Artificial <220> <223> N-terminal sequence-NP-C-terminal sequence (excluding reference aas) <400> 57 Ser Gly Pro Asn Pro Asn Pro Asn Pro Asn Pro Asn Ser Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met 20 25 30 Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr 35 40 45 Gly Ser Tyr Asn Pro Asn Pro Asn Pro Asn Pro Asn Pro Gly Ser 50 55 60 <210> 58 <211> 66 <212> PRT <213> Artificial <220> <223> N-terminal sequence-NP-C-terminal sequence (excluding reference aas) <400> 58 Ala Lys Val His Pro Asn Pro Asn Pro Asn Pro Asn Ser Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met 20 25 30 Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr 35 40 45 Gly Ser Tyr Asn Pro Asn Pro Asn Pro Asn Pro His Val Asp Val Trp 50 55 60 Gly Gln 65 <210> 59 <211> 61 <212> PRT <213> Artificial <220> <223> N-terminal sequence-NP-C-terminal sequence (excluding reference aas) <400> 59 Ala Ala Trp Asn Pro Asn Pro Asn Pro Asn Pro Asn Pro Asn Gly Gly 1 5 10 15 Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met Asp 20 25 30 Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr Gly 35 40 45 Ser Asn Pro Asn Pro Asn Pro Asn Pro Asn Pro Asn Val 50 55 60 <210> 60 <211> 66 <212> PRT <213> Artificial <220> <223> N-terminal sequence-NP-C-terminal sequence (excluding reference aas) <400> 60 Ala Lys Gly Ile Thr Gly Thr Lys Lys Tyr Gln Ser Ser Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met 20 25 30 Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr 35 40 45 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Ala Glu Tyr Phe Gln His Trp 50 55 60 Gly Gln 65 <210> 61 <211> 61 <212> PRT <213> Artificial <220> <223> N-terminal sequence-NP-C-terminal sequence (excluding reference aas) <400> 61 Ser Gly Ser Thr Tyr Ile Ser Asn Val Asn His Lys Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met Asp 20 25 30 Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr Gly 35 40 45 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Gly Gly Ser 50 55 60 <210> 62 <211> 59 <212> PRT <213> Artificial <220> <223> N-terminal sequence-NP-C-terminal sequence (excluding reference aas) <400> 62 Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Pro Asp Gly Gly Ser 1 5 10 15 Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met Asp Arg 20 25 30 Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr Gly Ser 35 40 45 Asp Arg Phe Ser Gly Ser Lys Ser Gly Gly Ser 50 55 <210> 63 <211> 61 <212> PRT <213> Artificial <220> <223> N-terminal sequence-NP-C-terminal sequence (excluding reference aas) <400> 63 Gly Val Pro Lys Glu Lys Glu Lys Glu Lys Val Ser Thr Ala Val Gly 1 5 10 15 Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met Asp Arg Ile 20 25 30 Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr Gly Ser Ala 35 40 45 Pro Leu Glu Val Pro Lys Glu Lys Glu Lys Glu Lys Val 50 55 60 <210> 64 <211> 56 <212> PRT <213> Artificial <220> <223> N-terminal sequence - NP - C-terminal sequence (excluding reference aas) <400> 64 Ser Val Val Val Thr Ser His Gln Ala Pro Gly Glu Gly Gly Ser Gly 1 5 10 15 Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met Asp Arg Ile 20 25 30 Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr Gly Gly Ser 35 40 45 Thr Pro Leu Lys Ser Leu Ala Ser 50 55 <210> 65 <211> 447 <212> PRT <213> Human <400> 65 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Leu Thr Gly Ala Glu Tyr Phe Gln His Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 <210> 66 <211> 217 <212> PRT <213> Human <400> 66 Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly 20 25 30 Tyr Asp Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu 35 40 45 Leu Ile Tyr Gly Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser Ser 85 90 95 Leu Ser Gly Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 110 Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu 115 120 125 Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe 130 135 140 Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val 145 150 155 160 Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys 165 170 175 Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser 180 185 190 His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu 195 200 205 Lys Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 67 <211> 499 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 67 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp Gly 100 105 110 Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Ile 115 120 125 Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr 130 135 140 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Asp Val Trp 145 150 155 160 Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 165 170 175 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 180 185 190 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 195 200 205 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 210 215 220 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 225 230 235 240 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 245 250 255 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 260 265 270 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 275 280 285 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 290 295 300 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 305 310 315 320 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 325 330 335 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 340 345 350 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 355 360 365 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 370 375 380 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 385 390 395 400 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 405 410 415 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 420 425 430 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 435 440 445 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 450 455 460 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 465 470 475 480 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 485 490 495 Ser Pro Gly <210> 68 <211> 499 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 68 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp Gly 100 105 110 Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met 115 120 125 Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr 130 135 140 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Asp Val Trp 145 150 155 160 Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 165 170 175 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 180 185 190 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 195 200 205 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 210 215 220 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 225 230 235 240 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 245 250 255 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 260 265 270 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 275 280 285 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 290 295 300 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 305 310 315 320 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 325 330 335 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 340 345 350 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 355 360 365 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 370 375 380 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 385 390 395 400 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 405 410 415 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 420 425 430 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 435 440 445 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 450 455 460 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 465 470 475 480 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 485 490 495 Ser Pro Gly <210> 69 <211> 504 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 69 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 50 55 60 Pro Asp Gly Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly 65 70 75 80 Gly Arg Met Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser 85 90 95 Phe Arg Tyr Gly Ser Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 100 105 110 Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser 115 120 125 Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg 130 135 140 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys Leu Thr Gly Ala Glu 145 150 155 160 Tyr Phe Gln His Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala 165 170 175 Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser 180 185 190 Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe 195 200 205 Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly 210 215 220 Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu 225 230 235 240 Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr 245 250 255 Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys 260 265 270 Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 275 280 285 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 290 295 300 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 305 310 315 320 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 325 330 335 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 340 345 350 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 355 360 365 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 370 375 380 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 385 390 395 400 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 405 410 415 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 420 425 430 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 435 440 445 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 450 455 460 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 465 470 475 480 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 485 490 495 Lys Ser Leu Ser Leu Ser Pro Gly 500 <210> 70 <211> 504 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 70 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Pro Asn Pro Asn Pro Asn Pro Asn Pro Asn Ser 50 55 60 Pro Asp Gly Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly 65 70 75 80 Gly Arg Met Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser 85 90 95 Phe Arg Tyr Gly Ser Tyr Asn Pro Asn Pro Asn Pro Asn Pro Asn Pro 100 105 110 Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser 115 120 125 Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg 130 135 140 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys Leu Thr Gly Ala Glu 145 150 155 160 Tyr Phe Gln His Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala 165 170 175 Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser 180 185 190 Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe 195 200 205 Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly 210 215 220 Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu 225 230 235 240 Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr 245 250 255 Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys 260 265 270 Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 275 280 285 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 290 295 300 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 305 310 315 320 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 325 330 335 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 340 345 350 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 355 360 365 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 370 375 380 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 385 390 395 400 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 405 410 415 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 420 425 430 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 435 440 445 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 450 455 460 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 465 470 475 480 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 485 490 495 Lys Ser Leu Ser Leu Ser Pro Gly 500 <210> 71 <211> 499 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 71 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Val His Pro Asn Pro Asn Pro Asn Pro Asn Ser Pro Asp Gly 100 105 110 Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met 115 120 125 Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr 130 135 140 Gly Ser Tyr Asn Pro Asn Pro Asn Pro Asn Pro His Val Asp Val Trp 145 150 155 160 Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 165 170 175 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 180 185 190 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 195 200 205 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 210 215 220 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 225 230 235 240 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 245 250 255 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 260 265 270 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 275 280 285 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 290 295 300 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 305 310 315 320 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 325 330 335 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 340 345 350 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 355 360 365 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 370 375 380 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 385 390 395 400 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 405 410 415 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 420 425 430 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 435 440 445 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 450 455 460 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 465 470 475 480 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 485 490 495 Ser Pro Gly <210> 72 <211> 504 <212> PRT <213> Artificial <220> <223> HC grafted with NP <400> 72 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Ser Gly 20 25 30 Ser Gly Ser Gly Ser Gly Ser Gly Ser Pro Asp Gly Gly Ser Gly Gly 35 40 45 Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met Asp Arg Ile Gly 50 55 60 Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr Gly Ser Tyr Gly 65 70 75 80 Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Met Ser Trp Val Arg Gln 85 90 95 Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ala Ile Ser Gly Ser Gly 100 105 110 Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser 115 120 125 Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg 130 135 140 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys Leu Thr Gly Ala Glu 145 150 155 160 Tyr Phe Gln His Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala 165 170 175 Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser 180 185 190 Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe 195 200 205 Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly 210 215 220 Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu 225 230 235 240 Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr 245 250 255 Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys 260 265 270 Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 275 280 285 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 290 295 300 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 305 310 315 320 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 325 330 335 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 340 345 350 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 355 360 365 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 370 375 380 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 385 390 395 400 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 405 410 415 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 420 425 430 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 435 440 445 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 450 455 460 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 465 470 475 480 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 485 490 495 Lys Ser Leu Ser Leu Ser Pro Gly 500 <210> 73 <211> 500 <212> PRT <213> Artificial <220> <223> NP-grafted HC[[ID=²6]] <400> 73 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Pro Asn Pro Asn Lys Asn Pro Asn Pro Asn Pro 50 55 60 Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met 65 70 75 80 It should be noted that in the translation, the "²6" in the original text seems to be an incorrect character. It is retained as is according to the requirements, but it may need to be corrected in the original source for a proper translation.Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr 85 90 95 Gly Ser Asn Pro Asn Glu Asn Pro Asn Pro Asn Pro Gly Ser Thr Tyr 100 105 110 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser 115 120 125 Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 130 135 140 Ala Val Tyr Tyr Cys Ala Lys Leu Thr Gly Ala Glu Tyr Phe Gln His 145 150 155 160 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly 165 170 175 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 180 185 190 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 195 200 205 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 210 215 220 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 225 230 235 240 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val 245 250 255 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys 260 265 270 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 275 280 285 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 290 295 300 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 305 310 315 320 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 325 330 335 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 340 345 350 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 355 360 365 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 370 375 380 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 385 390 395 400 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 405 410 415 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 420 425 430 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 435 440 445 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 450 455 460 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 465 470 475 480 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 485 490 495 Leu Ser Pro Gly 500 <210> 74 <211> 502 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 74 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Thr Tyr Ile Ser Asn Val Asn His Lys Pro 50 55 60 Asp Gly Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly 65 70 75 80 Arg Met Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe 85 90 95 Arg Tyr Gly Ser Asn Thr Lys Val Asp Lys Lys Val Glu Gly Gly Ser 100 105 110 Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp 115 120 125 Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu 130 135 140 Asp Thr Ala Val Tyr Tyr Cys Ala Lys Leu Thr Gly Ala Glu Tyr Phe 145 150 155 160 Gln His Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 165 170 175 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 180 185 190 Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 195 200 205 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 210 215 220 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 225 230 235 240 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 245 250 255 Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu 260 265 270 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 275 280 285 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 290 295 300 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 305 310 315 320 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 325 330 335 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 340 345 350 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 355 360 365 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 370 375 380 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 385 390 395 400 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 405 410 415 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 420 425 430 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 435 440 445 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 450 455 460 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 465 470 475 480 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 485 490 495 Leu Ser Leu Ser Pro Gly 500 <210> 75 <211> 500 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 75 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Pro Asp 50 55 60 Gly Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg 65 70 75 80 Met Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg 85 90 95 Tyr Gly Ser Asp Arg Phe Ser Gly Ser Lys Ser Gly Gly Ser Thr Tyr 100 105 110 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser 115 120 125 Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 130 135 140 Ala Val Tyr Tyr Cys Ala Lys Leu Thr Gly Ala Glu Tyr Phe Gln His 145 150 155 160 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly 165 170 175 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 180 185 190 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 195 200 205 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 210 215 220 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 225 230 235 240 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val 245 250 255 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys 260 265 270 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 275 280 285 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 290 295 300 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 305 310 315 320 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 325 330 335 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 340 345 350 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 355 360 365 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 370 375 380 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 385 390 395 400 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 405 410 415 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 420 425 430 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 435 440 445 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 450 455 460 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 465 470 475 480 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 485 490 495 Leu Ser Pro Gly 500 <210> 76 <211> 501 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 76 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Ser Gly 20 25 30 Ser Gly Ser Gly Ser Gly Ser Gly Ser Pro Asp Gly Gly Ser Gly Gly 35 40 45 Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met Asp Arg Ile Gly 50 55 60 Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr Gly Ser Tyr Gly 65 70 75 80 Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Met Ser Trp Val Arg Gln 85 90 95 Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ala Ile Ser Gly Ser Gly 100 105 110 Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser 115 120 125 Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg 130 135 140 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys Leu Thr Gly Ala Glu 145 150 155 160 Tyr Phe Gln His Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala 165 170 175 Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser 180 185 190 Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe 195 200 205 Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly 210 215 220 Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu 225 230 235 240 Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr 245 250 255 Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg 260 265 270 Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu 275 280 285 Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 290 295 300 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 305 310 315 320 Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly 325 330 335 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn 340 345 350 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 355 360 365 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro 370 375 380 Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 385 390 395 400 Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn 405 410 415 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 420 425 430 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 435 440 445 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg 450 455 460 Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys 465 470 475 480 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 485 490 495 Ser Leu Ser Leu Gly 500 <210> 77 <211> 493 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 77 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Ser Gly 20 25 30 Ser Gly Ser Gly Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe 35 40 45 Gly Gly Arg Met Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn 50 55 60 Ser Phe Arg Tyr Gly Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly Met 65 70 75 80 Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ala 85 90 95 Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly 100 105 110 Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln 115 120 125 Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys 130 135 140 Leu Thr Gly Ala Glu Tyr Phe Gln His Trp Gly Gln Gly Thr Leu Val 145 150 155 160 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 165 170 175 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 180 185 190 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 195 200 205 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 210 215 220 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 225 230 235 240 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 245 250 255 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 260 265 270 Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe 275 280 285 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 290 295 300 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 305 310 315 320 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 325 330 335 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 340 345 350 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 355 360 365 Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 370 375 380 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 385 390 395 400 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 405 410 415 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 420 425 430 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 435 440 445 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 450 455 460 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 465 470 475 480 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 485 490 <210> 78 <211> 497 <212> PRT <213> Artificial <220> <223> HC grafted with NP <400> 78 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Val Val Val Thr Ser His Gln Ala Pro Gly Glu Gly 50 55 60 Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met 65 7​​​​​​​​​​​​​​​​​Tyr Cys Ala Lys Leu Thr Gly Ala Glu Tyr Phe Gln His Trp Gly Gln 145 150 155 160 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 165 170 175 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 180 185 190 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 195 200 205 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 210 215 220 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 225 230 235 240 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 245 250 255 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 260 265 270 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 275 280 285 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 290 295 300 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 305 310 315 320 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 325 330 335 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 340 345 350 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 355 360 365 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 370 375 380 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 385 390 395 400 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 405 410 415 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 420 425 430 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 435 440 445 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 450 455 460 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 465 470 475 480 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 485 490 495 Gly <210> 79 <211> 499 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 79 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Ile Thr Gly Thr Lys Lys Tyr Gln Ser Ser Pro Asp Gly 100 105 110 Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly Arg Met 115 120 125 Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr 130 135 140 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Ala Glu Tyr Phe Gln His Trp 145 150 155 160 Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 165 170 175 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 180 185 190 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 195 200 205 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 210 215 220 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 225 230 235 240 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 245 250 255 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 260 265 270 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 275 280 285 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 290 295 300 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 305 310 315 320 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 325 330 335 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 340 345 350 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 355 360 365 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 370 375 380 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 385 390 395 400 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 405 410 415 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 420 425 430 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 435 440 445 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 450 455 460 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 465 470 475 480 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 485 490 495 Ser Pro Gly <210> 80 <211> 267 <212> PRT <213> Artificial <220> <223> LC grafted with NP <400> 80 Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly 20 25 30 Tyr Asp Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu 35 40 45 Leu Ile Tyr Gly Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asn Pro Asn 85 90 95 Pro Asn Pro Asn Pro Asn Pro Asn Gly Gly Ser Gly Gly Ser Leu Arg 100 105 110 Arg Ser Ser Cys Phe Gly Gly Arg Met Asp Arg Ile Gly Ala Gln Ser 115 120 125 Gly Leu Gly Cys Asn Ser Phe Arg Tyr Gly Ser Asn Pro Asn Pro Asn 130 135 140 Pro Asn Pro Asn Pro Asn Val Phe Gly Ser Gly Thr Lys Val Thr Val 145 150 155 160 Leu Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser 165 170 175 Ser Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser 180 185 190 Asp Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser 195 200 205 Pro Val Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn 210 215 220 Asn Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp 225 230 235 240 Lys Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr 245 250 255 Val Glu Lys Thr Val Ala Pro Thr Glu Cys Ser 260 265 <210> 81 <211> 274 <212> PRT <213> Artificial <220> <223> NP grafted LC <400> 81 Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly 20 25 30 Tyr Asp Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu 35 40 45 Leu Ile Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Pro 50 55 60 Asp Gly Gly Ser Gly Gly Ser Leu Arg Arg Ser Ser Cys Phe Gly Gly 65 70 75 80 Arg Met Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe 85 90 95 Arg Tyr Gly Ser Tyr Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly Asn 100 105 110 Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr 115 120 125 Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln Ala Glu Asp Glu Ala Asp 130 135 140 Tyr Tyr Cys Gln Ser Tyr Asp Ser Ser Leu Ser Gly Val Val Phe Gly 145 150 155 160 Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro Lys Ala Ala Pro Ser 165 170 175 Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu Gln Ala Asn Lys Ala 180 185 190 Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro Gly Ala Val Thr Val 195 200 205 Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala Gly Val Glu Thr Thr 210 215 220 Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala Ala Ser Ser Tyr Leu 225 230 235 240 Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg Ser Tyr Ser Cys Gln 245 250 255 Val Thr His Glu Gly Ser Thr Val Glu Lys Thr Val Ala Pro Thr Glu 260 265 270 Cys Ser <210> 82 <211> 366 <212> DNA <213> Artificial <220> <223> NP-grafted HC-encoding construct <400> 82 gaagtgcagc tgctggaaag cggcggaggc ctggtgcagc ctggcggatc tctgagactg 60 agctgtgccg ccagcggctt caccttcagc agctacgcca tgagctgggt gcgccaggcc 120 cctggaaaag gcctggaatg ggtgtccgcc atctctggca gcggcggcag cacctactac 180 gccgattctg tgaagggccg gttcaccatc agccgggaca acagcaagaa caccctgtac 240 ctgcagatga acagcctgcg ggccgaggac accgccgtgt actactgtac aagcgtgcac 300 caggaaacaa agaagtacca gagcagcccc gacggcggca gtggcggaag tctgagaaga 360 agctcc 366 <210> 83 <211> 525 <212> DNA <213> Artificial <220> <223> HC coding construct grafted with NP <400> 83 gaagttcagc tgctggaatc tggcggcgga ctggttcaac ctggcggatc tctgagactg 60 agctgtgccg ccagcggctt tacatttggc agcggctctg gatctggctc cggaagcgga 120 tctcctgatg gtggaagcgg aggcagcctg agaagaagca gctgtttcgg cggcagaatg 180 gacagaatcg gcgcccaatc tggcctgggc tgcaacagct ttagatacgg cagctacggc 240 tccggcagtg gttccggtag tggctctgga atgagctggg ttcgacaggc ccctggcaaa 300 ggccttgaat gggtgtccgc catttctggc agcggaggct ctacctacta cgccgatagc 360 gtgaagggca gattcaccat cagccgggac aacagcaaga acaccctgta cctgcagatg 420 aactccctga gagccgagga caccgccgtg tactattgcg ccaaactgac aggcgccgag 480 tacttccagc attggggaca gggaaccctg gtcacagtct cttca 525 <210> 84 <211> 333 <212> DNA <213> Human <400> 84 cagtctgtgc tgacacagcc tcctagtgtg tctggcgccc ctggccagag agtgaccatc 60 agctgtaccg gcagcagctc caacatcgga gccggctatg acgtgcactg gtatcagcag 120 ctgcctggca ccgcccccaa actgctgatc tacggcaaca gcaaccggcc cagcggcgtg 180 cccgatagat tttccggcag caagagcggc accagcgcca gcctggctat tactggactg 240 caggccgagg acgaggccga ctactactgc cagagctacg acagcagcct gagcggcgtg 300 gtgtttggcg gcggaacaaa gctgaccgtg cta 333 <210> 85 <211> 98 <212> PRT <213> Human <400> 85 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys <210> 86 <211> 17 <212> PRT <213> Human <400> 86 Ala Glu Tyr Phe Gln His Trp Gly Gln Gly Thr Leu Val Thr Val Ser 1 5 10 15 Ser <210> 87 <211> 329 <212> PRT <213> Human <400> 87 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105 110 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135 140 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 225 230 235 240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly 325 <210> 88 <211> 99 <212> PRT <213> Human <400> 88 Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly 20 25 30 Tyr Asp Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu 35 40 45 Leu Ile Tyr Gly Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser Ser 85 90 95 Leu Ser Gly <210> 89 <211> 12 <212> PRT <213> Human <400> 89 Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 1 5 10 <210> 90 <211> 106 <212> PRT <213> Human <400> 90 Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser 1 5 10 15 Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp 20 25 30 Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro 35 40 45 Val Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn 50 55 60 Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys 65 70 75 80 Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val 85 90 95 Glu Lys Thr Val Ala Pro Thr Glu Cys Ser 100 105 <210> 91 <211> 18 <212> PRT <213> Artificial <220> <223> NP variants <220> <221> misc_feature <222> (1)..(1) <223> Xaa can be any naturally occurring amino acid <220> <221> misc_feature <222> (5)..(7) <223> Xaa can be any naturally occurring amino acid <220> <221> misc_feature <222> (11)..(13) <223> Xaa can be any naturally occurring amino acid <220> <221> misc_feature <222> (15)..(15) <223> Xaa can be any naturally occurring amino acid <400> 91 Xaa Cys Phe Gly Xaa Xaa Xaa Asp Arg Ile Xaa Xaa Xaa Ser Xaa Leu 1 5 10 15 Gly Cys <210> 92 <211> 7 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 92 Ser Gly Phe Thr Phe Ser Ser 1 5 <210> 93 <211> 10 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 93 Ser Gly Phe Thr Phe Gly Ser Gly Ser Gly 1 5 10 <210> 94 <211> 6 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 94 Gly Ser Gly Ser Gly Met 1 5 <210> 95 <211> 13 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 95 Ser Gly Phe Thr Phe Gly Ser Gly Ser Gly Ser Gly Ser 1 5 10 <210> 96 <211> 9 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 96 Ser Gly Ser Gly Ser Gly Ser Gly Met 1 5 <210> 97 <211> 16 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 97 Ser Gly Phe Thr Phe Gly Ser Gly Ser Gly Ser Gly Gly Ser Gly Gly 1 5 10 15 <210> 98 <211> 12 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 98 Gly Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly Met 1 5 10 <210> 99 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 99 Ser Gly Phe Thr Phe Gly Ser Gly Ser Gly Ser Gly Ser Gly Gly Gly 1 5 10 15 Ser Gly Gly <210> 100 <211> 11 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 100 Gly Ser Gly Ser Gly Gly Ser Gly Ser Gly Met 1 5 10 <210> 101 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 101 Ser Pro Ala Val Val Tyr Ile Glu Ile Leu Asp Arg His Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 102 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 102 Gly Ser Gly Arg Glu Val Pro Ile Ser Asn Gly Ser Gly Phe Val Val 1 5 10 15 Ala Met <210> 103 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 103 Ser Gly Ala Val Val Tyr Ile Glu Ile Leu Asp Arg His Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 104 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 104 Ser Ser Ser Asp Arg Ser Ala Leu Leu Lys Ser Lys Leu Arg Ala Leu 1 5 10 15 Leu Thr Ala Pro Arg 20 <210> 105 <211> twenty four <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 105 Ser Gly Phe Thr Phe Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly 1 5 10 15 Ser Pro Asp Gly Gly Ser Gly Gly 20 <210> 106 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 106 Gly Ser Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Met 1 5 10 15 <210> 107 <211> 25 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 107 Ser Gly Phe Thr Phe Ser Ser Asp Arg Ser Ala Leu Leu Lys Ser Lys 1 5 10 15 Leu Arg Ala Leu Leu Thr Ala Pro Arg 20 25 <210> 108 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 108 Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Met 1 5 10 <210> 109 <211> 4 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 109 Ile Ser Gly Ser 1 <210> 110 <211> 4 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 110 Gly Gly Ser Thr 1 <210> 111 <211> 8 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 111 Ile Ser Gly Ser Gly Ser Gly Ser 1 5 <210> 112 <211> 8 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 112 Gly Ser Gly Ser Gly Gly Ser Thr 1 5 <210> 113 <211> 9 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 113 Ile Ser Gly Ser Gly Ser Gly Ser Gly 1 5 <210> 114 <211> 10 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 114 Gly Ser Ser Gly Ser Gly Ser Gly Ser Thr 1 5 10 <210> 115 <211> 11 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 115 Ile Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly 1 5 10 <210> 116 <211> 12 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 116 Gly Ser Ser Gly Ser Gly Ser Gly Ser Gly Ser Thr 1 5 10 <210> 117 <211> 12 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 117 Ile Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Gly 1 5 10 <210> 118 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 118 Gly Glu Lys Glu Lys Glu Lys Val Ser Thr Ala Val Gly Ser Thr 1 5 10 15 <210> 119 <211> 13 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 119 Ile Ser Gly Ser Ala Val Val Asn Gly Gly Ser Gly Gly 1 5 10 <210> 120 <211> 9 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 120 Gly Lys Ile Ala Ile Gly Gly Ser Thr 1 5 <210> 121 <211> 14 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 121 Ile Ser Gly Pro Asn Pro Asn Lys Asn Pro Asn Pro Gly Gly 1 5 10 <210> 122 <211> 13 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 122 Gly Ser Asn Glu Asn Pro Asn Pro Asn Pro Gly Ser Thr 1 5 10 <210> 123 <211> 15 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 123 Ile Ser Gly Ser Val Val Val Thr Ser His Gly Gly Ser Gly Gly 1 5 10 15 <210> 124 <211> 10 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 124 Gly Gly Ser Gly Ser Gly Ser Gly Ser Thr 1 5 10 <210> 125 <211> 15 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 125 Ile Ser Gly Ser Ala Val Val Asn Val Arg Gly Gly Ser Gly Gly 1 5 10 15 <210> 126 <211> 11 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 126 Gly Gly Asp Lys Ile Ala Ile Gly Gly Ser Thr 1 5 10 <210> 127 <211> 15 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 127 Ile Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Ser Gly Gly 1 5 10 15 <210> 128 <211> 15 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 128 Ile Ser Gly Leu Ala Val Gln Ile Arg Arg Gly Gly Ser Gly Gly 1 5 10 15 <210> 129 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 129 Gly Gly Ser Gly Arg Glu Thr Leu Thr Leu Tyr Val Gly Ser Thr 1 5 10 15 <210> 130 <211> 16 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 130 Ile Ser Gly Ser Ala Val Val Asn Val Arg Ala Gly Gly Ser Gly Gly 1 5 10 15 <210> 131 <211> 16 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 131 Ile Ser Gly Ser Tyr Ala Met Ser Trp Val Arg Gly Gly Ser Gly Gly 1 5 10 15 <210> 132 <211> 13 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 132 Gly Ser Tyr Ala Met Ser Trp Val Arg Gln Gly Ser Thr 1 5 10 <210> 133 <211> 16 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 133 Ile Ser Gly Pro Asn Pro Asn Lys Asn Pro Asn Pro Asn Pro Gly Gly 1 5 10 15 <210> 134 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 134 Gly Ser Asn Pro Asn Glu Asn Pro Asn Pro Asn Pro Gly Ser Thr 1 5 10 15 <210> 135 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 135 Ile Ser Gly Ser Ala Val Val Asn Val Arg Ala Asp Gly Gly Ser Gly 1 5 10 15 Gly <210> 136 <211> 12 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 136 Gly Ser Gly Asp Lys Ile Ala Ile Gly Gly Ser Thr 1 5 10 <210> 137 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 137 Ile Ser Gly Ser Gly Ser Gly Ser Gly Ser Pro Asp Gly Gly Ser Gly 1 5 10 15 Gly <210> 138 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 138 Ile Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Gly Ser Gly 1 5 10 15 Gly <210> 139 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 139 Ile Ser Gly Ser Val Val Val Thr Ser His Gln Ala Pro Gly Ser Gly 1 5 10 15 Gly <210> 140 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 140 Gly Ser Gly Glu Lys Lys Lys Leu Lys Ser Leu Ala Tyr Gly Ser Thr 1 5 10 15 <210> 141 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 141 Ile Ser Gly Arg Tyr Asn Ile Leu Lys Ile Gln Lys Val Gly Ser Gly 1 5 10 15 Gly <210> 142 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 142 Gly Gly Ser Gly Glu Tyr Leu Ile Thr Tyr Gln Ile Met Gly Ser Thr 1 5 10 15 <210> 143 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 143 Ile Ser Gly Arg Gln Leu Leu Phe Cys Arg Val Thr Leu Gly Ser Gly 1 5 10 15 Gly <210> 144 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 144 Gly Gly Ser Gly Glu Gln Ala Tyr Pro Glu Tyr Leu Ile Thr Tyr Gly 1 5 10 15 Ser Thr <210> 145 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 145 Ile Ser Val Val Val Thr Ser His Gln Ala Pro Gly Glu Gly Gly Ser 1 5 10 15 Gly Gly <210> 146 <211> 12 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 146 Gly Glu Lys Lys Lys Leu Lys Ser Leu Ala Ser Thr 1 5 10 <210> 147 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 147 Ile Ser Gly Val Val Thr Ser His Gln Ala Pro Gly Glu Gly Gly Ser 1 5 10 15 Gly Gly <210> 148 <211> 12 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 148 Gly Glu Lys Lys Lys Leu Lys Ser Leu Gly Ser Thr 1 5 10 <210> 149 <211> 12 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 149 Gly Glu Lys Lys Lys Leu Lys Ser Gly Gly Ser Thr 1 5 10 <210> 150 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 150 Ile Ser Gly Ser Val Thr Ser His Gln Ala Pro Gly Glu Gly Gly Ser 1 5 10 15 Gly Gly <210> 151 <211> 12 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 151 Gly Glu Lys Lys Lys Gly Lys Ser Gly Gly Ser Thr 1 5 10 <210> 152 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 152 Ile Ser Val Val Val Thr Ser His Gln Ala Pro Gly Ser Gly Gly Ser 1 5 10 15 Gly Gly <210> 153 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 153 Ile Ser Val Val Val Thr Ser His Gln Ala Pro Thr Ser Gly Gly Ser 1 5 10 15 Gly Gly <210> 154 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 154 Ile Ser Val Val Val Thr Ser His Gln Ser Pro Thr Pro Gly Gly Ser 1 5 10 15 Gly Gly <210> 155 <211> 12 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 155 Gly Gly Ser Thr Pro Leu Lys Ser Leu Ala Ser Thr 1 5 10 <210> 156 <211> 12 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 156 Gly Ser Thr Pro Lys Leu Lys Ser Leu Ala Ser Thr 1 5 10 <210> 157 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 157 Ile Ser Val Val Val Thr Ser His Pro Thr Pro Gly Glu Gly Gly Ser 1 5 10 15 Gly Gly <210> 158 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 158 Ile Ser Val Val Val Thr Ser His Gln Ala Pro Ser Pro Gly Ser Thr 1 5 10 15 Gly Gly <210> 159 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 159 Ile Ser Val Val Val Thr Ser His Gln Ala Asn Gly Ser Gly Gly Ser 1 5 10 15 Gly Gly <210> 160 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 160 Ile Ser Gly Ser Ala Val Val Asn Val Arg Ala Pro Asp Gly Gly Ser 1 5 10 15 Gly Gly <210> 161 <211> 13 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 161 Gly Ser Lys Gly Asp Lys Ile Ala Ile Gly Gly Ser Thr 1 5 10 <210> 162 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 162 Ile Ser Thr Ser Ala Ser Leu Ala Ile Thr Gly Pro Asp Gly Gly Ser 1 5 10 15 Gly Gly <210> 163 <211> 13 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 163 Gly Ser Asp Arg Phe Ser Gly Ser Lys Ser Gly Ser Thr 1 5 10 <210> 164 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 164 Ile Ser Gly Phe Ile Leu Pro Ile Glu Val Tyr Pro Asp Gly Gly Ser 1 5 10 15 Gly Gly <210> 165 <211> 13 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 165 Gly Ser Lys Val Arg Phe Asp Tyr Asp Leu Phe Ser Thr 1 5 10 <210> 166 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 166 Ile Ser Ser Ala Leu Leu Lys Ser Lys Leu Arg Ala Leu Leu Thr Ala 1 5 10 15 Pro Gly <210> 167 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 167 Gly Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Thr 1 5 10 <210> 168 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 168 Ile Ser Gly Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Gly Gly Ser 1 5 10 15 Gly Gly <210> 169 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 169 Gly Ser Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Gly Ser Thr 1 5 10 15 <210> 170 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 170 Ile Ser Gly Pro Asn Pro Asn Lys Asn Pro Asn Pro Asn Pro Gly Ser 1 5 10 15 Gly Gly <210> 171 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 171 Ile His Pro Leu Gln Asn Arg Trp Ala Leu Trp Phe Phe Lys Gly Ser 1 5 10 15 Gly Gly <210> 172 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 172 Gly Gly Ser Gly Asn Leu Arg Leu Ile Ser Lys Phe Asp Thr Val Thr 1 5 10 15 <210> 173 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 173 Ile Ser Gly Ser Val Thr Ile Phe Ser Leu Ala Thr Asn Glu Gly Ser 1 5 10 15 Gly Gly <210> 174 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 174 Gly Gly Ser Gly Lys Thr Thr Trp His Arg Ile Ser Val Phe Gly Gly 1 5 10 15 Ser Thr <210> 175 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 175 Ile Tyr Leu Glu Gly Lys Ile Asp Tyr Gly Glu Tyr Met Asp Gly Ser 1 5 10 15 Gly Gly <210> 176 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 176 Gly Gly Ser Asn Val Arg Arg Gln Ala Thr Thr Ile Ile Ala Asp Asn 1 5 10 15 Ile Thr <210> 177 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 177 Ile Ser Gly Ser Val Gln Gly Ile Ile Asn Phe Glu Gln Lys Gly Ser 1 5 10 15 Gly Gly <210> 178 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 178 Gly Gly Ser Gly Pro Val Lys Val Trp Gly Ser Ile Lys Gly Gly Gly 1 5 10 15 Ser Thr <210> 179 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 179 Ile Ser Gly Val Val Val Thr Ser His Gln Ala Pro Gly Glu Gly Gly 1 5 10 15 Ser Gly Gly <210> 180 <211> 13 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 180 Gly Glu Lys Lys Lys Leu Lys Ser Leu Ala Gly Ser Thr 1 5 10 <210> 181 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 181 Ile Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 182 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 182 Gly Ser Asp Arg Phe Ser Gly Ser Lys Ser Gly Gly Ser Thr 1 5 10 <210> 183 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 183 Ile Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 184 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 184 Gly Ser Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Thr 1 5 10 <210> 185 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 185 Ile Ser Gly Thr Tyr Ile Ser Asn Val Asn His Lys Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 186 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 186 Gly Ser Asn Thr Lys Val Asp Lys Lys Val Glu Gly Ser Thr 1 5 10 <210> 187 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 187 Ile Ser Gly Gly Phe Ile Leu Pro Ile Glu Val Tyr Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 188 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 188 Gly Ser Lys Val Arg Phe Asp Tyr Asp Leu Phe Gly Ser Thr 1 5 10 <210> 189 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 189 Ile Ser Gly Ser Val Val Val Thr Ser His Gln Ala Pro Gly Gly Gly 1 5 10 15 Ser Gly Gly <210> 190 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 190 Gly Glu Lys Lys Lys Leu Lys Ser Leu Ala Tyr Gly Ser Thr 1 5 10 <210> 191 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 191 Gly Glu Lys Pro Lys Pro Lys Pro Leu Ala Tyr Gly Ser Thr 1 5 10 <210> 192 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 192 Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Gly Ser Thr 1 5 10 <210> 193 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 193 Ile Ser Gly Pro Asn Pro Asn Lys Asn Pro Asn Pro Asn Pro Gly Gly 1 5 10 15 Ser Gly Gly <210> 194 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 194 Ile Ser Gly Asp Ile Tyr Leu Ala Ile Asn Ile Thr Asn Gly Glu Gly 1 5 10 15 Ser Gly Gly <210> 195 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 195 Gly Gly Ser Gly Asp Ile Tyr Leu Ala Ile Asn Ile Thr Asn Gly Glu 1 5 10 15 Ser Thr <210> 196 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 196 Ile Ser Gly Ser Ala Thr Lys Ala Val Ser Val Leu Lys Gly Asp Gly 1 5 10 15 Ser Gly Gly <210> 197 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 197 Gly Gly Ser Gly Val Gln Gly Ile Ile Asn Phe Glu Gln Lys Gly Gly 1 5 10 15 Ser Thr <210> 198 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 198 Ile Ser Gly Ser Val Pro Lys Glu Lys Glu Lys Glu Lys Val Ser Thr 1 5 10 15 Ala Val Gly Gly 20 <210> 199 <211> 13 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 199 Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Thr 1 5 10 <210> 200 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 200 Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Thr 1 5 10 15 <210> 201 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 201 Ile Ser Gly Ser Ser Gly Ala Val Val Asn Val Arg Ala Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 202 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 202 Gly Ser Lys Gly Asp Lys Ile Ala Ile Trp Thr Thr Gly Ser Thr 1 5 10 15 <210> 203 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 203 Ile Ser Gly Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 204 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 204 Ile Ser Gly Ser Thr Ser Ala Ser Leu Ala Ile Thr Gly Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 205 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 205 Gly Ser Asp Arg Phe Ser Gly Ser Lys Ser Gly Gly Gly Ser Thr 1 5 10 15 <210> 206 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 206 Ile Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 207 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 207 Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Ser Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 208 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 208 Gly Ser Tyr Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Thr 1 5 10 15 <210> 209 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 209 Ile Ser Thr Gln Thr Tyr Ile Ser Asn Val Asn His Lys Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 210 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 210 Gly Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Thr 1 5 10 15 <210> 211 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 211 Ile Ser Gly Ser Thr Tyr Ile Ser Asn Val Asn His Lys Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 212 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 212 Gly Ser Asn Thr Lys Val Asp Lys Lys Val Glu Gly Gly Ser Thr 1 5 10 15 <210> 213 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 213 Ile Ser Gly Pro Asn Pro Asn Pro Asn Pro Asn Pro Asn Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 214 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 214 Gly Ser Asn Pro Asn Pro Asn Pro Asn Pro Asn Pro Gly Ser Thr 1 5 10 15 <210> 215 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 215 Ile Ser Ala Val Gln Val Lys Leu Glu Leu Gly His Arg Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 216 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 216 Gly Ser Asn His Leu Arg Ser Glu Lys Leu Thr Phe Asn Ser Thr 1 5 10 15 <210> 217 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 217 Ile Ser Gly Phe Ile Leu Pro Ile Glu Val Tyr Phe Lys Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 218 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 218 Gly Ser Pro Arg Lys Val Arg Phe Asp Tyr Asp Leu Phe Ser Thr 1 5 10 15 <210> 219 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 219 Ile Ser Gly Ser Gly Phe Ile Leu Pro Ile Glu Val Tyr Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 220 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 220 Gly Ser Lys Val Arg Phe Asp Tyr Asp Leu Phe Gly Gly Ser Thr 1 5 10 15 <210> 221 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 221 Gly Ser Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Thr 1 5 10 15 <210> 222 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 222 Ile Ser Asp Arg Ser Ala Leu Leu Lys Ser Lys Leu Arg Ala Leu Leu 1 5 10 15 Thr Ala Pro Arg 20 <210> 223 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 223 Gly Ser Asp Arg Ser Ala Leu Leu Lys Ser Lys Leu Arg Ala Ser Thr 1 5 10 15 <210> 224 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 224 Ile Ser Asp Arg Ser Ala Leu Leu Lys Ser Lys Leu Arg Ala Leu Leu 1 5 10 15 Thr Ala Pro Gly 20 <210> 225 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 225 Gly Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Thr 1 5 10 15 <210> 226 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 226 Ile Ser Gly Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 227 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 227 Gly Ser Lys Thr His Thr Ser Pro Pro Ser Pro Gly Gly Ser Thr 1 5 10 15 <210> 228 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 228 Ile Ser Gly Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 229 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 229 Gly Ser Tyr Ser Ser Tyr Ala Met Ser Trp Val Arg Gly Gly Ser Thr 1 5 10 15 <210> 230 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 230 Gly Ser Tyr Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Gly Ser Thr 1 5 10 15 <210> 231 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 231 Ile Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 232 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 232 Ile Ser Gly Ser Pro Asn Pro Asn Pro Asn Pro Asn Pro Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 233 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 233 Gly Ser Tyr Pro Asn Pro Asn Pro Asn Pro Asn Pro Ser Gly Ser Thr 1 5 10 15 <210> 234 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 234 Ile Ser Gly Pro Asn Pro Asn Lys Asn Pro Asn Pro Asn Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 235 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 235 Gly Ser Tyr Asn Pro Asn Glu Asn Pro Asn Pro Asn Pro Gly Ser Thr 1 5 10 15 <210> 236 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 236 Ile Ser Gly Pro Asn Pro Asn Pro Asn Pro Asn Pro Asn Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 237 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 237 Gly Ser Tyr Asn Pro Asn Pro Asn Pro Asn Pro Asn Pro Gly Ser Thr 1 5 10 15 <210> 238 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 238 Ile Ser Gly Ser Val Val Val Thr Ser His Gln Ala Pro Gly Gly Ser 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 239 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 239 Ile Ser Gly Ser Val Val Tyr Ile Glu Ile Leu Asp Arg His Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 240 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 240 Gly Ser Gly Arg Glu Val Pro Ile Ser Asn Gly Ser Gly Gly Ser Thr 1 5 10 15 <210> 241 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 241 Ile Ser Gly Ala Val Val Tyr Ile Glu Ile Leu Asp Arg His Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 242 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 242 Ile Ser Pro Ala Val Val Tyr Ile Glu Ile Leu Asp Arg His Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 243 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 243 Gly Ser Gly Arg Glu Val Pro Ile Ser Asn Gly Ser Gly Phe Ser Thr 1 5 10 15 <210> 244 <211> 11 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 244 Cys Ala Lys Ser Pro Asp Gly Gly Ser Gly Gly 1 5 10 <210> 245 <211> 4 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 245 Gly Ser Tyr Gly 1 <210> 246 <211> 9 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 246 Gly Ser Tyr Gln His Trp Gly Gln Gly 1 5 <210> 247 <211> 13 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 247 Cys Ala Lys Val His Gln Glu Thr Gly Gly Ser Gly Gly 1 5 10 <210> 248 <211> 11 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 248 Gly Ser Trp His Val Gln His Trp Gly Gln Gly 1 5 10 <210> 249 <211> 15 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 249 Cys Ala Lys Val His Gln Glu Thr Pro Asp Gly Gly Ser Gly Gly 1 5 10 15 <210> 250 <211> 13 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 250 Gly Ser Tyr Glu Trp His Val Gln His Trp Gly Gln Gly 1 5 10 <210> 251 <211> 16 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 251 Cys Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Gly Gly 1 5 10 15 <210> 252 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 252 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Asp Val Trp 1 5 10 15 Gly Gln Gly <210> 253 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 253 Cys Thr Ser Val His Gln Glu Thr Ser Ser Pro Asp Gly Gly Ser Gly 1 5 10 15 Gly <210> 254 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 254 Gly Ser Tyr Ser Tyr Glu Trp His Val Asp Val Trp Gly Gln Gly 1 5 10 15 <210> 255 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 255 Cys Ala Lys Thr His Thr Ser Pro Pro Ser Pro Asp Gly Gly Ser Gly 1 5 10 15 Gly <210> 256 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 256 Gly Ser Ser Pro Pro Ser Pro Tyr Phe Gln His Trp Gly Gln Gly 1 5 10 15 <210> 257 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 257 Cys Thr Ser Val His Gln Glu Thr Lys Ser Ser Pro Asp Gly Gly Ser 1 5 10 15 Gly Gly <210> 258 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 258 Gly Ser Tyr Ser Asn Tyr Glu Trp His Val Asp Val Trp Gly Gln Gly 1 5 10 15 <210> 259 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 259 Cys Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp 1 5 10 15 Gly Gly <210> 260 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 260 Cys Thr Ser Val His Gln Glu Thr Lys Lys Ser Ser Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 261 <211> 17 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 261 Gly Ser Tyr Ser Tyr Asn Tyr Glu Trp His Val Asp Val Trp Gly Gln 1 5 10 15 Gly <210> 262 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 262 Cys Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Gly Gly 1 5 10 15 Ser Gly Gly <210> 263 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 263 Cys Thr Ser Val His Gln Glu Thr Lys Lys Gln Ser Ser Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 264 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 264 Gly Ser Tyr Ser Tyr Tyr Asn Tyr Glu Trp His Val Asp Val Trp Gly 1 5 10 15 Gln Gly <210> 265 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 265 Cys Ala Lys Val His Pro Asn Pro Asn Pro Asn Pro Asn Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 266 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 266 Gly Ser Asn Pro Asn Pro Asn Pro Asn Pro His Val Asp Val Trp Gly 1 5 10 15 Gln Gly <210> 267 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 267 Cys Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly 20 <210> 268 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 268 Cys Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 269 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 269 Cys Ala Lys Leu Thr Val Val Val Thr Ser His Gln Ala Pro Gly Glu 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 270 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 270 Gly Glu Lys Lys Lys Leu Lys Ser Leu Ala Tyr Phe Gln His Trp Gly 1 5 10 15 Gln Gly <210> 271 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 271 Cys Ala Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 272 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 272 Gly Ser Lys Thr His Thr Ser Pro Pro Ser Pro Tyr Phe Gln His Trp 1 5 10 15 Gly Gln Gly <210> 273 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 273 Cys Ala Lys Val Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 274 <211> 17 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 274 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Val Gln His Trp Gly Gln 1 5 10 15 Gly <210> 275 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 275 Cys Ala Lys Val His Thr Lys Lys Tyr Gln Ser Ser Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 276 <211> 17 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 276 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr His Val Gln His Trp Gly Gln 1 5 10 15 Gly <210> 277 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 277 Cys Ala Lys Leu Thr Val Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 278 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 278 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Gln His Trp 1 5 10 15 Gly Gln Gly <210> 279 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 279 Cys Ala Lys Leu Thr Ala Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 280 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 280 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Asn Tyr Phe Gln His Trp 1 5 10 15 Gly Gln Gly <210> 281 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 281 Cys Ala Lys Gly Ile Thr Gly Thr Lys Lys Tyr Gln Ser Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 282 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 282 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Ala Glu Tyr Phe Gln His Trp 1 5 10 15 Gly Gln Gly <210> 283 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 283 Gly Ser Tyr Asp Tyr Val Trp Gly Ser Tyr Ala Tyr Phe Gln His Trp 1 5 10 15 Gly Gln Gly <210> 284 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 284 Cys Ala Lys Leu Thr Ser Val Val Val Thr Ser His Gln Ala Pro Gly 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 285 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 285 Gly Glu Lys Lys Lys Leu Lys Ser Leu Ala Tyr Tyr Phe Gln His Trp 1 5 10 15 Gly Gln Gly <210> 286 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 286 Cys Ala Lys Val His Pro Asn Pro Asn Pro Asn Pro Asn Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 287 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 287 Gly Ser Tyr Asn Pro Asn Pro Asn Pro Asn Pro His Val Asp Val Trp 1 5 10 15 Gly Gln Gly <210> 288 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 288 Cys Ala Lys Leu Thr Gln Val Lys Leu Glu Leu Gly His Arg Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 289 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 289 Gly Ser Asn His Leu Arg Ser Glu Lys Leu Thr Tyr Phe Gln His Trp 1 5 10 15 Gly Gln Gly <210> 290 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 290 Cys Ala Lys Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 291 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 291 Cys Ala Lys Thr Gln Thr Tyr Ile Ser Asn Val Asn His Lys Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 292 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 292 Gly Ser Asn Thr Lys Val Asp Lys Lys Ala Glu Tyr Phe Gln His Trp 1 5 10 15 Gly Gln Gly <210> 293 <211> 20 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 293 Gly Ser Tyr Ser Tyr Thr Thr Tyr Asn Tyr Glu Trp His Val Asp Val 1 5 10 15 Trp Gly Gln Gly 20 <210> 294 <211> twenty two <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 294 Cys Ala Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro 1 5 10 15 Asp Gly Gly Ser Gly Gly 20 <210> 295 <211> 20 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 295 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Asp Val His 1 5 10 15 Trp Gly Gln Gly 20 <210> 296 <211> twenty three <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 296 Cys Ala Lys Leu Thr Ala Glu Glu Trp Lys Lys Lys Tyr Glu Lys Glu 1 5 10 15 Lys Glu Lys Asn Lys Gly Ser 20 <210> 297 <211> twenty one <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 297 Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ala Glu Tyr Phe Gln 1 5 10 15 His Trp Gly Gln Gly 20 <210> 298 <211> twenty three <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 298 Cys Ala Asp Ser Ser Asp Arg Ser Ala Leu Leu Lys Ser Lys Leu Arg 1 5 10 15 Ala Leu Leu Thr Ala Pro Arg 20 <210> 299 <211> twenty one <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 299 Gly Ser Asn His Leu Arg Ser Glu Lys Leu Thr Phe Asn Tyr Phe Gln 1 5 10 15 His Trp Gly Gln Gly 20 <210> 300 <211> twenty three <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 300 Cys Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Tyr Gln Ser Ser 1 5 10 15 Pro Asp Gly Gly Ser Gly Gly 20 <210> 301 <211> twenty one <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 301 Gly Ser Tyr Ser Tyr Thr Tyr Thr Tyr Asn Tyr Glu Trp His Val Asp 1 5 10 15 Val Trp Gly Gln Gly 20 <210> 302 <211> twenty three <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 302 Cys Ala Lys Leu Thr Asp Arg Ser Ala Leu Leu Lys Ser Lys Leu Arg 1 5 10 15 Ala Leu Leu Thr Ala Pro Arg 20 <210> 303 <211> twenty three <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 303 Cys Ala Lys Leu Thr Ala Val Gln Val Lys Leu Glu Leu Gly His Arg 1 5 10 15 Pro Asp Gly Gly Ser Gly Gly 20 <210> 304 <211> 25 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 304 Cys Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Tyr Gln 1 5 10 15 Ser Ser Pro Asp Gly Gly Ser Gly Gly 20 25 <210> 305 <211> twenty three <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 305 Gly Ser Tyr Ser Tyr Thr Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His 1 5 10 15 Val Asp Val Trp Gly Gln Gly 20 <210> 306 <211> 25 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 306 Cys Ala Lys Leu Thr Ala Val Gln Val Lys Leu Glu Leu Gly His Arg 1 5 10 15 Ala Gln Pro Asp Gly Gly Ser Gly Gly 20 25 <210> 307 <211> twenty three <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 307 Gly Ser Pro Val Asn His Leu Arg Ser Glu Lys Leu Thr Phe Asn Tyr 1 5 10 15 Phe Gln His Trp Gly Gln Gly 20 <210> 308 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 308 Ser Ser Ser Asn Ile Gly Ser Lys Leu Arg Ala Leu Leu Thr Ala Pro 1 5 10 15 Arg <210> 309 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 309 Gly Ser Gly Ser Gly Gly Ser Gly Gly Ser Gly Ser Gly Tyr Asp 1 5 10 15 <210> 310 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 310 Gly Ser Tyr Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly Asp 1 5 10 <210> 311 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 311 Ser Ser Leu Gly Gln Ile Gln Leu Thr Ile Arg His Ser Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 312 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 312 Gly Ser Asn Lys Leu Ile Val Val Val His Ala Ser Arg Asn Leu Ile 1 5 10 15 Gly Tyr Asp <210> 313 <211> twenty two <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 313 Ser Pro Leu Gly Gln Ile Gln Leu Thr Ile Arg His Ser Ser Gln Pro 1 5 10 15 Asp Gly Gly Ser Gly Gly 20 <210> 314 <211> 20 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 314 Gly Ser Arg Asn Lys Leu Ile Val Val Val His Ala Ser Arg Asn Leu 1 5 10 15 Ile Ala Tyr Asp 20 <210> 315 <211> twenty two <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 315 Ser Ser Leu Gly Gln Ile Gln Leu Thr Ile Arg His Ser Ser Gln Pro 1 5 10 15 Asp Gly Gly Ser Gly Gly 20 <210> 316 <211> 20 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 316 Gly Ser Arg Asn Lys Leu Ile Val Val Val His Ala Ser Arg Asn Leu 1 5 10 15 Ile Gly Tyr Asp 20 <210> 317 <211> twenty two <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 317 Ser Ser Ser Asn Ile Gly Ser Ala Leu Leu Lys Ser Lys Leu Arg Ala 1 5 10 15 Leu Leu Thr Ala Pro Arg 20 <210> 318 <211> 20 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 318 Gly Ser Ser Asp Arg Ser Ala Leu Leu Lys Ser Lys Leu Arg Ala Leu 1 5 10 15 Leu Thr Ala Asp 20 <210> 319 <211> 20 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 319 Gly Ser Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Ser Gly 1 5 10 15 Ser Gly Tyr Asp 20 <210> 320 <211> twenty four <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 320 Ser Ser Ser Asn Ile Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly 1 5 10 15 Ser Pro Asp Gly Gly Ser Gly Gly 20 <210> 321 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 321 Gly Ser Tyr Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly Tyr Asp 1 5 10 15 <210> 322 <211> 7 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 322 Asn Ser Asn Arg Pro Ser Gly 1 5 <210> 323 <211> 6 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 323 Tyr Gly Gly Ser Gly Ser 1 5 <210> 324 <211> 11 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 324 Gly Ser Gly Ser Asn Ser Asn Arg Pro Ser Gly 1 5 10 <210> 325 <211> 13 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 325 Tyr Gly Lys Thr His Thr Ser Pro Pro Ser Pro Gly Gly 1 5 10 <210> 326 <211> 17 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 326 Gly Gly Lys Thr His Thr Ser Pro Pro Ser Pro Gly Asn Arg Pro Ser 1 5 10 15 Gly <210> 327 <211> 14 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 327 Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Gly Gly 1 5 10 <210> 328 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 328 Gly Gly Lys Thr His Thr Ser Pro Pro Ser Pro Ser Gly Asn Arg Pro 1 5 10 15 Ser Gly <210> 329 <211> 14 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 329 Tyr Gly Ser Lys Thr His Thr Ser Pro Pro Ser Pro Gly Gly 1 5 10 <210> 330 <211> 15 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 330 Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Gly Gly Ser Gly Gly 1 5 10 15 <210> 331 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 331 Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly Asn Arg Pro Ser Gly 1 5 10 15 <210> 332 <211> 16 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 332 Tyr Gly Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser Pro Gly Gly 1 5 10 15 <210> 333 <211> 20 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 333 Gly Gly Ser Gly Ser Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly Asn 1 5 10 15 Arg Pro Ser Gly 20 <210> 334 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 334 Tyr Thr Ser Ala Ser Leu Ala Ile Thr Gly Pro Asp Gly Gly Ser Gly 1 5 10 15 Gly <210> 335 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 335 Gly Ser Asp Arg Phe Ser Gly Ser Lys Ser Gly Asn Arg Pro Ser Gly 1 5 10 15 <210> 336 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 336 Tyr Gly Phe Ile Leu Pro Ile Glu Val Tyr Pro Asp Gly Gly Ser Gly 1 5 10 15 Gly <210> 337 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 337 Gly Ser Lys Val Arg Phe Asp Tyr Asp Leu Phe Asn Arg Pro Ser Gly 1 5 10 15 <210> 338 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 338 Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Gly Gly Ser Gly 1 5 10 15 Gly <210> 339 <211> 17 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 339 Gly Ser Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly Asn Arg Pro Ser 1 5 10 15 Gly <210> 340 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 340 Tyr Gly Val Pro Lys Glu Lys Glu Lys Glu Lys Val Ser Thr Ala Val 1 5 10 15 Gly Gly <210> 341 <211> 17 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 341 Gly Ser Ala Pro Leu Glu Val Pro Lys Glu Lys Glu Lys Glu Lys Val 1 5 10 15 Gly <210> 342 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 342 Tyr Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Pro Asp Gly Gly Ser 1 5 10 15 Gly Gly <210> 343 <211> 17 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 343 Gly Ser Asp Arg Phe Ser Gly Ser Lys Ser Gly Gly Asn Arg Pro Ser 1 5 10 15 Gly <210> 344 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 344 Tyr Gly Gly Ser Gly Ser Gly Ser Gly Ser Gly Pro Asp Gly Gly Ser 1 5 10 15 Gly Gly <210> 345 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 345 Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly 1 5 10 15 Gly Gly <210> 346 <211> 17 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 346 Gly Ser Tyr Glu Lys Glu Lys Glu Lys Asn Lys Thr Leu Lys Asn Val 1 5 10 15 Gly <210> 347 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 347 Tyr Gly Thr Tyr Ile Ser Asn Val Asn His Lys Pro Asp Gly Gly Ser 1 5 10 15 Gly Gly <210> 348 <211> 17 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 348 Gly Ser Asn Thr Lys Val Asp Lys Lys Val Glu Gly Asn Arg Pro Ser 1 5 10 15 Gly <210> 349 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 349 Tyr Gly Ala Glu Glu Trp Lys Lys Lys Tyr Glu Lys Glu Lys Glu Lys 1 5 10 15 Gly Gly <210> 350 <211> 17 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 350 Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 1 5 10 15 Gly <210> 351 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 351 Tyr Gly Gly Phe Ile Leu Pro Ile Glu Val Tyr Pro Asp Gly Gly Ser 1 5 10 15 Gly Gly <210> 352 <211> 17 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 352 Gly Ser Lys Val Arg Phe Asp Tyr Asp Leu Phe Gly Asn Arg Pro Ser 1 5 10 15 Gly <210> 353 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 353 Tyr Gly Ser Ala Leu Leu Lys Ser Lys Leu Arg Ala Leu Leu Thr Ala 1 5 10 15 Pro Gly <210> 354 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 354 Gly Ser Asp Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly Asn Arg Pro 1 5 10 15 Ser Gly <210> 355 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 355 Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 356 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 356 Tyr Gly Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 357 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 357 Gly Ser Asp Arg Phe Ser Gly Ser Lys Ser Gly Gly Gly Asn Arg Pro 1 5 10 15 Ser Gly <210> 358 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 358 Tyr Gly Gly Thr Tyr Ile Ser Asn Val Asn His Lys Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 359 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 359 Gly Ser Asn Thr Lys Val Asp Lys Lys Val Glu Gly Gly Asn Arg Pro 1 5 10 15 Ser Gly <210> 360 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 360 Tyr Thr Gln Thr Tyr Ile Ser Asn Val Asn His Lys Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 361 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 361 Gly Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Asn Arg Pro 1 5 10 15 Ser Gly <210> 362 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 362 Tyr Gly Gly Gly Phe Ile Leu Pro Ile Glu Val Tyr Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 363 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 363 Gly Ser Lys Val Arg Phe Asp Tyr Asp Leu Phe Gly Gly Asn Arg Pro 1 5 10 15 Ser Gly <210> 364 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 364 Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 365 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 365 Gly Ser Tyr Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly Asn Arg Pro 1 5 10 15 Ser Gly <210> 366 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 366 Tyr Gly Ser Gln Val Lys Leu Glu Leu Gly His Arg Ala Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 367 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 367 Gly Ser Val Asn His Leu Arg Ser Glu Lys Leu Thr Ser Gly Asn Arg 1 5 10 15 Pro Ser Gly <210> 368 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 368 Tyr Pro Ala Val Val Tyr Ile Glu Ile Leu Asp Arg His Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 369 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 369 Gly Ser Gly Arg Glu Val Pro Ile Ser Asn Gly Ser Gly Phe Asn Arg 1 5 10 15 Pro Ser Gly <210> 370 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 370 Tyr Gly Gly Val Val Tyr Ile Glu Ile Leu Asp Arg His Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 371 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 371 Gly Ser Gly Arg Glu Val Pro Ile Ser Asn Gly Ser Gly Gly Asn Arg 1 5 10 15 Pro Ser Gly <210> 372 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 372 Tyr Gly Ala Val Val Tyr Ile Glu Ile Leu Asp Arg His Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 373 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 373 Tyr Gly Ser Arg Ser Ala Leu Leu Lys Ser Lys Leu Arg Ala Leu Leu 1 5 10 15 Thr Ala Pro Arg 20 <210> 374 <211> 20 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 374 Gly Ser Asp Gly Ser Gly Ser Gly Gly Ser Gly Ser Gly Ser Gly Asn 1 5 10 15 Arg Pro Ser Gly 20 <210> 375 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 375 Tyr Gly Ser Arg Ser Ala Leu Leu Lys Ser Lys Leu Arg Ala Leu Leu 1 5 10 15 Thr Ala Pro Gly 20 <210> 376 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 376 Tyr Gly Ser Ala Val Gln Val Lys Leu Glu Leu Gly His Arg Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 377 <211> 20 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 377 Gly Ser Asn His Leu Arg Ser Glu Lys Leu Thr Phe Asn Ser Gly Asn 1 5 10 15 Arg Pro Ser Gly 20 <210> 378 <211> 15 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 378 Cys Gly Ser Gly Ser Gly Ser Gly Pro Asp Gly Gly Ser Gly Gly 1 5 10 15 <210> 379 <211> 10 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 379 Gly Ser Gly Ser Gly Ser Gly Ser Gly Phe 1 5 10 <210> 380 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 380 Cys Gln Ser Tyr Asp Ile Leu Pro Ile Glu Pro Asp Gly Gly Ser Gly 1 5 10 15 Gly <210> 381 <211> 11 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 381 Gly Ser Arg Phe Asp Tyr Asp Gly Val Val Phe 1 5 10 <210> 382 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 382 Cys Gly Ser Gly Ser Gly Ser Gly Ser Gly Pro Asp Gly Gly Ser Gly 1 5 10 15 Gly <210> 383 <211> 12 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 383 Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Phe 1 5 10 <210> 384 <211> 17 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 384 Cys Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Gly Gly Ser Gly 1 5 10 15 Gly <210> 385 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 385 Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Phe 1 5 10 <210> 386 <211> 18 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 386 Cys Gln Ser Tyr Asp Lys Leu Glu Leu Gly His Pro Asp Gly Gly Ser 1 5 10 15 Gly Gly <210> 387 <211> 12 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 387 Gly Ser His Leu Arg Ser Glu Lys Gly Val Val Phe 1 5 10 <210> 388 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 388 Cys Gln Ser Tyr Asp Ile Leu Pro Ile Glu Val Tyr Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 389 <211> 13 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 389 Gly Ser Lys Val Arg Phe Asp Tyr Asp Gly Val Val Phe 1 5 10 <210> 390 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 390 Cys Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Pro Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 391 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 391 Cys Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Asp Gly Gly 1 5 10 15 Ser Gly Gly <210> 392 <211> 19 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 392 Cys Gln Ser Tyr Asp Gly Phe Ile Leu Pro Ile Glu Val Tyr Gly Gly 1 5 10 15 Ser Gly Gly <210> 393 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 393 Gly Ser Lys Val Arg Phe Asp Tyr Asp Leu Phe Gly Val Val Phe 1 5 10 15 <210> 394 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 394 Cys Gln Ser Tyr Asp Lys Leu Glu Leu Gly His Arg Ala Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 395 <211> 14 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 395 Gly Ser Val Asn His Leu Arg Ser Glu Lys Gly Val Val Phe 1 5 10 <210> 396 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 396 Cys Gln Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 397 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 397 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Val Phe 1 5 10 15 <210> 398 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 398 Cys Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 399 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 399 Gly Ser Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Phe 1 5 10 15 <210> 400 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 400 Cys Gln Ser Tyr Asp Pro Asn Pro Asn Pro Asn Pro Asn Pro Asp Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 401 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 401 Gly Ser Asn Pro Asn Pro Asn Pro Asn Pro Ser Gly Val Val Phe 1 5 10 15 <210> 402 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 402 Cys Ala Ala Trp Asn Pro Asn Pro Asn Pro Asn Pro Asn Pro Asn Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 403 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 403 Gly Ser Asn Pro Asn Pro Asn Pro Asn Pro Asn Pro Asn Val Phe 1 5 10 15 <210> 404 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 404 Cys Gln Ser Tyr Asp Gln Val Lys Leu Glu Leu Gly His Arg Ala Gly 1 5 10 15 Gly Ser Gly Gly 20 <210> 405 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 405 Gly Ser Val Asn His Leu Arg Ser Glu Lys Leu Thr Gly Val Val Phe 1 5 10 15 <210> 406 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 406 Cys Gln Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 407 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 407 Cys Gln Ser Tyr Asp Gln Val Lys Leu Glu Leu Gly His Arg Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 408 <211> 15 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 408 Gly Ser Asn His Leu Arg Ser Glu Lys Leu Thr Gly Val Val Phe 1 5 10 15 <210> 409 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 409 Cys Gln Ser Tyr Asp Gly Phe Ile Leu Pro Ile Glu Val Tyr Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 410 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 410 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Asp Val Phe 1 5 10 15 <210> 411 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 411 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Val Val Phe 1 5 10 15 <210> 412 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 412 Cys Gly Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly 20 <210> 413 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 413 Gly Ser Tyr Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Gly Phe 1 5 10 15 <210> 414 <211> twenty two <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 414 Cys Gln Ser Tyr Asp Ser Ser Ala Leu Leu Lys Ser Lys Leu Arg Ala 1 5 10 15 Leu Leu Thr Ala Pro Arg 20 <210> 415 <211> twenty two <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 415 Cys Gln Ser Ala Val Gln Val Lys Leu Glu Leu Gly His Arg Ala Pro 1 5 10 15 Asp Gly Gly Ser Gly Gly 20 <210> 416 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 416 Gly Ser Val Asn His Leu Arg Ser Glu Lys Leu Thr Phe Asn Val Phe 1 5 10 15 <210> 417 <211> twenty two <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 417 Cys Gln Ser Tyr Asp Gln Val Lys Leu Glu Leu Gly His Arg Ala Pro 1 5 10 15 Asp Gly Gly Ser Gly Gly 20 <210> 418 <211> twenty two <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 418 Cys Gln Ser Tyr Asp Gly Phe Ile Leu Pro Ile Glu Val Tyr Phe Pro 1 5 10 15 Asp Gly Gly Ser Gly Gly 20 <210> 419 <211> 16 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 419 Gly Ser Arg Lys Val Arg Phe Asp Tyr Asp Leu Phe Gly Val Val Phe 1 5 10 15 <210> 420 <211> twenty two <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 420 Cys Gln Ser Tyr Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro 1 5 10 15 Asp Gly Gly Ser Gly Gly 20 <210> 421 <211> 17 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 421 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Gly Val Val 1 5 10 15 Phe <210> 422 <211> twenty three <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 422 Cys Gln Ser Tyr Asp Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser 1 5 10 15 Pro Asp Gly Gly Ser Gly Gly 20 <210> 423 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 423 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Ser Gly Val 1 5 10 15 Val Phe <210> 424 <211> twenty four <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 424 Cys Gln Ser Tyr Asp Ala Val Gln Val Lys Leu Glu Leu Gly His Arg 1 5 10 15 Ala Pro Asp Gly Gly Ser Gly Gly 20 <210> 425 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 425 Gly Ser Val Asn His Leu Arg Ser Glu Lys Leu Thr Phe Asn Gly Val 1 5 10 15 Val Phe <210> 426 <211> 25 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 426 Cys Gln Ser Tyr Asp Ser Ser Asp Arg Ser Ala Leu Leu Lys Ser Lys 1 5 10 15 Leu Arg Ala Leu Leu Thr Ala Pro Arg 20 25 <210> 427 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 427 Gly Ser Gly Gly Ser Val Asn His Leu Arg Ser Glu Lys Leu Thr Gly 1 5 10 15 Val Val Phe <210> 428 <211> twenty one <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 428 Gly Ser Asp Arg Ser Ala Leu Leu Lys Ser Lys Leu Arg Ala Leu Leu 1 5 10 15 Thr Ala Val Val Phe 20 <210> 429 <211> 25 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 429 Cys Gln Ser Tyr Asp Ser Ser Asp Arg Ser Ala Leu Leu Lys Ser Lys 1 5 10 15 Leu Arg Ala Leu Leu Thr Ala Pro Glu 20 25 <210> 430 <211> twenty three <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 430 Cys Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly Ser Gly 20 <210> 431 <211> 20 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 431 Gly Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Asp Val 1 5 10 15 Trp Gly Gln Gly 20 <210> 432 <211> 18 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 432 Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Asp Val Trp Gly 1 5 10 15 Gln Gly <210> 433 <211> twenty two <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 433 Cys Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp 1 5 10 15 Gly Gly Ser Gly Gly Ser 20 <210> 434 <211> twenty four <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 434 Gly Ser Gly Gly Tyr Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp 1 5 10 15 His Val Asp Val Trp Gly Gln Gly 20 <210> 435 <211> twenty one <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 435 Cys Gln Ser Tyr Asp Gln Val Lys Leu Glu Leu Gly His Arg Ala Gly 1 5 10 15 Gly Ser Gly Gly Ser 20 <210> 436 <211> twenty one <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 436 Gly Ser Gly Gly Ser Gly Ser Val Asn His Leu Arg Ser Glu Lys Leu 1 5 10 15 Thr Gly Val Val Phe 20 <210> 437 <211> twenty four <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 437 Gly Ser Gly Gly Ser Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp 1 5 10 15 His Val Asp Val Trp Gly Gln Gly 20 <210> 438 <211> twenty three <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 438 Gly Gly Gly Ser Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His 1 5 10 15 Val Asp Val Trp Gly Gln Gly 20 <210> 439 <211> twenty two <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 439 Cys Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro Tyr 1 5 10 15 Lys Gly Ala Asn Lys Lys 20 <210> 440 <211> 20 <212> PRT <213> Artificial <220> <223> N-terminal sequence (including reference aa) <400> 440 Ile Ser Gly Ser Val Val Val Thr Ser His Gln Ala Pro Gly Gly Gly 1 5 10 15 Ser Gly Gly Ser 20 <210> 441 <211> 19 <212> PRT <213> Artificial <220> <223> C-terminal sequence (including reference aa) <400> 441 Gly Ser Gly Gly Ser Gly Glu Lys Lys Lys Leu Lys Ser Leu Ala Tyr 1 5 10 15 Gly Ser Thr <210> 442 <211> 499 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 442 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Ser Val His Gln Glu Thr Lys Lys Tyr Gln Ser Ser Pro Asp Gly 100 105 110 Gly Ser Gly Gly Met Val Gln Gly Ser Gly Cys Phe Gly Arg Lys Met 115 120 125 Asp Arg Ile Ser Ser Ser Ser Gly Leu Gly Cys Lys Val Leu Arg Arg 130 135 140 Gly Ser Tyr Ser Tyr Thr Tyr Asn Tyr Glu Trp His Val Asp Val Trp 145 150 155 160 Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 165 170 175 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 180 185 190 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 195 200 205 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 210 215 220 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 225 230 235 240 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 245 250 255 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 260 265 270 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 275 280 285 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 290 295 300 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 305 310 315 320 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 325 330 335 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 340 345 350 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 355 360 365 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 370 375 380 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 385 390 395 400 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 405 410 415 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 420 425 430 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 435 440 445 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 450 455 460 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 465 470 475 480 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 485 490 495 Ser Pro Gly <210> 443 <211> 500 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 443 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys...

Claims

1. An antibody or fragment thereof, comprising at least one heterologous amino acid sequence, wherein the at least one heterologous amino acid sequence consists of an N-terminal linker sequence (Ntls), a C-type natriuretic peptide (CNP) and a C-terminal linker sequence (Ctls), wherein the amino acid sequences of the heavy chain and the light chain of the antibody are selected from the following groups: The heavy chain is SEQ ID NO 445, and the light chain is SEQ ID NO 66; The heavy chain is SEQ ID NO 445, and the light chain is SEQ ID NO 447; The heavy chain is SEQ ID NO 445, and the light chain differs from SEQ ID NO 66 by LC_A33Y; and The antibody fragment is selected from Fab, Fab', Fab'-SH, F(ab')2 and Fv fragments; diabodies; single domain antibodies (Dab); linear antibodies; single chain antibody molecules (ScFv); and disulfide bond stabilized Fv antibody fragments (dsFv).

2. The antibody or fragment thereof according to claim 1, wherein the antibody or fragment thereof is a human or humanized antibody or fragment thereof.

3. The antibody or fragment thereof according to claim 2, wherein the antibody or fragment thereof is of the IgG class.

4. A composition comprising a compound of the antibody or fragment thereof according to any one of claims 1 to 3 and optionally a pharmaceutically acceptable carrier.

5. A nucleic acid or a mixture of nucleic acids encoding the antibody or fragment thereof according to any one of claims 1 to 3.

6. A host cell comprising the nucleic acid or nucleic acid mixture according to claim 5.

7. A method for producing an antibody or a fragment thereof, comprising culturing the host cell according to claim 6 under conditions suitable for expressing the antibody or the fragment thereof.

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