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

Description

TECHNICAL FIELD

[0001] 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 the fragment thereof, wherein the at least one heterologous amino acid sequence comprises an N-terminal linker sequence (Ntls), atrial natriuretic peptide (ANP), and a C-terminal linker sequence (Ctls). Optionally, at least a part 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 the following: when the heterologous amino acid sequence is incorporated into CDRH1, between the amino acid residue HC (heavy chain) res25 according to Kabat and the first amino acid residue of ANP; when the heterologous amino acid sequence is incorporated into CDRH2, between the amino acid residue HC res51 according to Kabat and the first amino acid residue of ANP; when the heterologous amino acid sequence is incorporated into CDRH3, between the amino acid sequence residue HC res92 according to Kabat and the first amino acid residue of ANP; when the heterologous amino acid sequence is incorporated into CDRL1, between the amino acid sequence residue LC (light chain) res26 according to Kabat and the first amino acid residue of ANP; when the heterologous amino acid sequence is incorporated into CDRL2, between the amino acid residue LC res49 according to Kabat and the first amino acid residue of ANP; and / or when the heterologous amino acid sequence is incorporated into CDRL3, between the amino acid residue LC res88 according to Kabat and the first amino acid residue of ANP. In addition, there are at least 9 amino acid residues between the last amino acid residue of ANP and the following: when the heterologous amino acid sequence is incorporated into CDRH1, the amino acid residue HC res35a according to Kabat; when the heterologous amino acid sequence is incorporated into CDRH2, the amino acid sequence residue HC res57 according to Kabat; when the heterologous amino acid sequence is incorporated into CDRH3, the amino acid residue HC res106 according to Kabat; when the heterologous amino acid sequence is incorporated into CDRL1, the amino acid residue LC res 32 according to Kabat; when the heterologous amino acid sequence is incorporated into CDRL2, the amino acid residue LC res57 according to Kabat; and / or when the heterologous amino acid sequence is incorporated into CDRL3, the amino acid residue LC res98 according to Kabat. The present invention also relates to the use of such an antibody or a fragment thereof in a therapeutic method, a composition comprising such an antibody or a fragment thereof, a nucleic acid or a nucleic acid mixture encoding such an antibody or a fragment thereof, a host cell comprising such a nucleic acid or such a nucleic acid mixture, and a method for producing such an antibody or a fragment thereof. BACKGROUND OF THE INVENTION

[0002] The natriuretic peptides are a family of three structurally related peptides with neurohumoral actions. Atrial natriuretic peptide (ANP) is a 28-amino acid peptide containing a central ring structure formed by a disulfide bond bridge between cysteine residues 7 and 23. Human ANP is expressed in atrial myocytes as a 153-amino acid long preprohormone. Signal peptide cleavage generates the prohormone form, which is subsequently further cleaved into mature ANP and an N-terminal residue, called 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 mainly produced and released by cardiomyocytes in the left and right atria, while BNP is mainly 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. The natriuretic peptides are encoded by three separate 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 the 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 effective natriuretic peptides that activate or bind to cell receptors and can elicit related physiological effects.

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

[0004] The cardiac hormones ANP and BNP are excreted upon ventricular and atrial stretch, for example due to hypervolemia. They exert a vasodilatory effect by relaxing vascular smooth muscle and result in a decrease in blood pressure. In the kidney, ANP causes an increase in urine excretion (diuresis) and an increase in the sodium ion concentration in 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 etal.Intensive Care Med Exp. 2014 Dec;2(1):28; Werner et al., Basic ResCardiol. 2016 Mar;111(2):22; Kimura et al., Respir Res. 2016 Feb 19;17:19). Activation of NPR-B by CNP 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 physiological effects of natriuretic peptides and their receptors make them attractive targets in 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 under 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 major obstacle to the therapeutic use of natriuretic peptides is their very short plasma half-life in organisms, 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 the endocytosis of the NPR-C receptor, natriuretic peptides are also effectively proteolytically degraded by neprilysin (NEP) and insulin-degrading enzyme (IDE). The related short-term biological effects of the administered natriuretic peptides limit their therapeutic use mainly to acute indications. For example, in different countries, the infusion of recombinant carperitide (ANP) and nesiritide (BNP) is approved for the treatment of acute decompensated heart failure.

[0007] The treatment of chronic diseases will be greatly facilitated by providing NPR-A and NPR-B agonists with increased plasma half-life, improved proteolytic stability, and extended action time.

[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), ulinastatin (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 I): P38). Other ANP and CNP derivatives are disclosed in US 9,193,777 and EP2432489 A, respectively.

[0009] In addition, natriuretic peptide fusions (including Fc fusions, albumin fusions) and pegylated natriuretic peptides have been described. For example, natriuretic peptide-Fc fusions are disclosed in US 2010 / 0310561, WO2008 / 154226, WO2010 / 117760, WO2006 / 107124, WO2008 / 136611 and WO2008 / 079995. Natriuretic peptide-albumin fusions are disclosed in US 7,521,424 and US 2014 / 0148390, and pegylated natriuretic peptides are disclosed in US 2014 / 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 mimetic grafted antibody library obtained by replacing the entire original CDRH3 region of the 2G12 antibody with an ANP mimetic peptide flanked by two random amino acid residues on both sides. Neither WO2005 / 060642 nor WO2005 / 082004 discloses any specific natriuretic peptide grafted antibody, let alone functionally characterizing these antibodies.

[0011] Object of the Invention

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

[0013] The above object is achieved by the teachings of the present independent claims. The inventors have surprisingly found that bioactive natriuretic peptide variants with significantly increased stability in serum compared to naturally occurring wild-type natriuretic peptides can be obtained by incorporating a natriuretic peptide amino acid sequence into one of the CDR regions of an immunoglobulin molecule or a fragment thereof, although the CDR regions of immunoglobulins are short in length and highly sequence-conserved, which poses a considerable conformational restraint on the incorporation of bioactive peptides. 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 to produce bioactive natriuretic peptide variants is the number of amino acid residues between the N-terminal and C-terminal flanks of the incorporated natriuretic peptide and the nearest neighbor CDR framework to which the incorporated natriuretic peptide is linked. When the number of N-terminal and C-terminal flanking amino acid residues between the natriuretic peptide and the adjacent CDR framework link is below 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 levels of protein fragmentation.

[0014] Accordingly, in a first aspect, the present invention relates to an antibody or a fragment thereof, said antibody or fragment thereof comprising at least one heterologous amino acid sequence incorporated into at least one CDR region of said antibody or fragment thereof, wherein said 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 part of said at least one CDR region is replaced by said at least one heterologous amino acid sequence incorporated therein, wherein

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

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

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

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

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

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

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

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

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

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

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

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

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

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

[0029] In other aspects, the present invention relates to such an antibody or its fragment for use in a therapeutic method, a composition comprising such an antibody or its fragment, a nucleic acid or nucleic acid mixture encoding such an antibody or its fragment, a host cell comprising such a nucleic acid or such a nucleic acid mixture, and a method for producing such an antibody or its fragment. Detailed Description

[0030] The present invention can be more readily understood by reference to the following detailed description of the invention and the examples contained therein.

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

[0032] The present inventors have discovered that bioactive natriuretic peptide variants with significantly increased stability in serum compared to naturally occurring wild-type natriuretic peptides can be obtained by incorporating a natriuretic peptide amino acid sequence into a CDR region of an immunoglobulin molecule or a fragment thereof. This discovery was completely unexpected. As is well known in the art, the relatively short length and high sequence conservation of immunoglobulin CDR regions (particularly evident in CDRL1, CDRL2, CDRL3, CDRH1, and CDRH2) impose considerable conformational constraints on the incorporation of bioactive peptides, and the surrounding immunoglobulin sequences may have an adverse effect on the expression, folding, and / or bioactivity of the incorporated peptide. In fact, the present inventors have found that the activity of natriuretic peptides incorporated within the immunoglobulin CDR regions varies widely, depending on the exact manner in which the natriuretic peptide-grafted antibodies are constructed. The decisive factor for the successful incorporation of a functional (i.e., bioactive) natriuretic peptide variant has proven to be the number of amino acid residues between the N-terminal and C-terminal flanks of the incorporated natriuretic peptide and the CDR framework to which the incorporated natriuretic peptide is nearest neighbor. When the N-terminal and C-terminal flanking amino acid residues between the natriuretic peptide and the adjacent CDR framework connection are below a certain number, no bioactive natriuretic peptide immunoglobulin fusion constructs are obtained.

[0033] The terms “incorporated,” “inserted,” “integrated,” “grafted,” and “embedded,” and “incorporating,” “inserting,” “integrating,” “grafting,” and “embedding” are used interchangeably herein. In the context of the present invention, these terms refer to the generation of a hybrid polynucleotide or polypeptide by introducing a heterologous sequence into the original sequence of an antibody or antibody fragment. Such incorporation can be carried out in any manner. Typically, an antibody or a fragment thereof containing a natriuretic peptide flanked by N-terminal and C-terminal linker sequences is produced by recombinant DNA technology and expressed as described herein.

[0034] Incorporating a natriuretic peptide flanked by N-terminal and C-terminal linker sequences into the CDR region of an original antibody or antibody fragment sequence may result in the deletion of at least a portion of the CDR region. For example, a nucleic acid sequence encoding the heterologous amino acid sequence comprising an N-terminal linker sequence, a natriuretic peptide, and a C-terminal linker sequence can 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 a heterologous amino acid sequence containing a 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 initial “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 a modified recombinant antibody molecule containing amino acid sequences of different origins.

[0036] The term "natriuretic peptide" refers to a peptide that induces natriuresis (i.e., sodium excretion by the kidney). 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 can have any origin. Natriuretic peptides include native natriuretic peptides (such as wild-type natriuretic peptides) and their mutant forms, as well as homologous natriuretic peptides from different species. However, the term also includes engineered natriuretic peptides, such as engineered chimeric variants of different natriuretic peptides. It is known that codon usage is different between species. Thus, when expressing a heterologous protein in a target cell, it may be necessary or at least beneficial to engineer the nucleic acid sequence to adapt to the codon usage of the target cell. Methods for designing and constructing derivatives of a given protein are well known to any person of ordinary skill in the art.

[0037] In a specific embodiment, the natriuretic peptide is selected from wild-type natriuretic peptides of any species and functional variants 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 native peptides and engineered peptides, that differ in their amino acid sequence and / or in the nucleic acid sequence encoding the amino acid sequence of a given natriuretic peptide (such as 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 functions of a naturally occurring natriuretic peptide (specifically, a wild-type natriuretic peptide). Specifically, "functional activity" means that the natriuretic peptide variant is able to bind to its respective receptor. In the case of the 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 a specific embodiment, the functional natriuretic peptide variant is able to perform at least about 50%, specifically at least about 60%, at least about 70%, at least about 80%, most specifically at least about 90% of one or more biological functions of a given natriuretic peptide (such as a wild-type natriuretic peptide of any given species), wherein the one or more biological functions include, but are not limited to, the binding of the natriuretic peptide to its respective receptor and / or the induction of an increase in intracellular cGMP.

[0039] The functional activity of a 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 fluorescence assay described in Example 3 50Values below 500 nM, more specifically below 250 nM, more specifically below 150 nM, more specifically below 100 nM, more specifically below 50 nM, and most specifically below 25 nM, then the (non-grafted) natriuretic peptide variant is considered to have functional activity.

[0040] Incorporating such a functional natriuretic peptide variant into a CDR region of an immunoglobulin molecule or a fragment thereof as described herein results in a natriuretic peptide-grafted immunoglobulin having natriuretic peptide functional activity, and the natriuretic peptide-grafted immunoglobulin has a significantly increased stability in serum 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 of measuring an increase in (intracellular) cyclic guanosine monophosphate (cGMP) directly or indirectly by assessing changes in cGMP-regulated cellular processes, then the natriuretic peptide-grafted immunoglobulin is considered to have biological activity (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 the natriuretic peptide-grafted immunoglobulin, significance is generally evaluated based on: i) comparison with a negative sample, such as an empty immunoglobulin scaffold, e.g., construct #209 (antibody comprising SEQ ID NO65 and SEQ ID NO 66, TPP-5657), ii) comparison with a positive sample, e.g., construct #117 (antibody comprising SEQID NO 67 and SEQ ID NO 66, TPP-5661), and iii) dose-dependence.

[0041] Although the functional natriuretic peptide variants of the present invention may contain any number of mutations including additions, deletions, and / or substitutions of one or more amino acids compared to a 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 ring 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 a specific embodiment, the functional natriuretic peptide variant has 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 regions of an immunoglobulin or its fragment as described herein. Specifically, the inventors have found that the discovery of the minimum requirements for satisfactory bioactivity of the grafted natriuretic peptide and the 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 human ANP having the sequence SEQ ID NO: 23, human BNP having the sequence SEQ ID NO: 24, human CNP having the sequence SEQ ID NO: 25, and a 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. Similarly, a natriuretic peptide having a sequence that deviates from the wild-type human natriuretic peptide ANP, BNP, and CNP sequences can have any natural source, such as a mutant version of the wild-type human natriuretic peptide, a homolog from a different species, or a modified natriuretic peptide. Methods for designing and constructing peptide variants are known to any person of ordinary skill in the art.

[0049] In a specific such embodiment, 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 a reference polynucleotide or polypeptide sequence, "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, respectively, in the reference polynucleotide or polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve maximum percent sequence identity. Conservative substitutions are not considered part of sequence identity. In a particular embodiment, any gaps introduced in the candidate sequence and / or the reference sequence total no more than 50%, 40%, 30%, 25%, 20%, 15% or 10% of the total number of residues in the reference sequence. In a particular embodiment, percent sequence identity is determined without introducing any gaps into the candidate sequence or the reference sequence (i.e., using a gapless alignment). Alignments for determining percent amino acid sequence identity can be achieved by a variety of methods well known to those of skill in the art, e.g., using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those of skill in the art can determine appropriate parameters for aligning sequences, including any algorithms required to achieve maximal alignment over the full length of the sequences being compared.

[0051] A natriuretic peptide having a given percent sequence identity to a given reference natriuretic peptide (e.g., human ANP having the amino acid sequence of SEQ ID NO: 23) may contain one or more mutations including one or more amino acid additions, deletions, and / or substitutions compared to the reference natriuretic peptide. According to the teachings of the present invention, the amino acids of said deletions, additions, and / or substitutions may be contiguous amino acids, or may be dispersed over the length of the amino acid sequence of the natriuretic peptide having a given percent sequence identity to the reference natriuretic peptide (e.g., human ANP having the amino acid sequence of SEQ ID NO: 23). At the DNA level, nucleic acid sequences encoding natriuretic peptides having a given percent sequence identity to 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 so long as the specified amino acid sequence identity to the reference natriuretic peptide is maintained. In a particular embodiment, the specified amino acid sequence identity is maintained and the natriuretic peptide variant is biologically active, i.e., a functional natriuretic peptide variant. Preferably, compared to the reference natriuretic peptide measured in the above assays, the reduction in biological activity of a natriuretic peptide having a given percent sequence identity to a given reference natriuretic peptide (e.g., human ANP having the amino acid sequence of SEQ ID NO: 23) is less than 90%, less than 80%, less than 70%, less than 60%, less than 50%, less than 25% or less than 10%.

[0053] As used herein, the term "antibody" means an immunoglobulin molecule, particularly a dimeric immunoglobulin molecule composed of four polypeptide chains: two heavy (H) chains and two light (L) chains, which are usually linked to each other 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 hypervariable regions, called complementarity determining regions (CDRs), between which there are more conserved regions, called framework regions (FRs). Each VH and VL typically consists of three CDRs and four FRs arranged from the amino terminus to the carboxy 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 the variable domain of an antibody that are necessary for antigen binding. Each variable domain typically has three CDR regions, designated CDR1, CDR2, and CDR3. Each complementarity determining region can include the amino acid residues in the "complementarity determining region" as defined by Kabat (e.g., with respect to the light chain variable domain, approximately residues 24 - 34 (CDRL1), 50 - 56 (CDRL2), and 89 - 97 (CDRL3), and with respect to the heavy chain variable domain, 31 - 35 (CDRH1), 50 - 65 (CDRH2), and 95 - 102 (CDRH3); (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, 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 some cases, the complementarity determining region can include amino acids from the CDR regions as defined by Kabat and the hypervariable loops.

[0055] Based on the amino acid sequence of their heavy chain constant domains, intact antibodies can be assigned to different "classes". There are five main classes of intact 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 immunoglobulin molecules of any main class (including IgG, IgE, IgM, IgD, IgA, and IgY) and immunoglobulin molecules of any subclass (including IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) isolated from nature or prepared by recombinant methods, and includes all conventionally known antibodies. A preferred class of immunoglobulin 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 the same active binding conformation as that present in natural antibodies, such that the binding of the observed target antigen to these chimeric proteins remains unchanged compared to the binding activity of the natural 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 contains at least one CDR region. Specifically, the antibody fragments of the present invention retain the ability to increase the serum half-life of a bioactive peptide incorporated therein (preferably a natriuretic peptide). 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 containing 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 remove the intermolecular disulfide bond interactions that exist between the CH1 and CL domains. The antibody fragments of the present invention can include the variable region alone or can include the variable region and all or part of the following: hinge region, CH1, CH2, CH3, and CL domains. Also included are antibody fragments that contain any combination of the variable region with the hinge region, CH1, CH2, CH3, and CL domains.

[0058] The antibody or its fragment constitutes 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 region of an antibody described herein may be increased compared to the naturally occurring natriuretic peptide.

[0059] Primarily, a heterologous amino acid sequence containing a natriuretic peptide can be incorporated into any immunoglobulin molecule or fragment thereof. Specifically, immunoglobulins of any species (including but not limited to humans, cows, mice, rats, pigs, dogs, sharks, llamas, and camels) and any main class and subclass can be used according to the present invention. However, for therapeutic use, human or humanized antibodies may be preferred. In the context of the present invention, the term "human antibody" refers to an antibody having a human immunoglobulin amino acid sequence, including antibodies isolated from a human immunoglobulin library, isolated from human B cells, or isolated from transgenic animals expressing 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 the 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), the antibody being based on human germline sequences; or (ii) a chimeric antibody or fragment thereof, wherein 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, wherein the CDRs of the variable domain are from a non-human source and one or more frameworks of the variable domain are from a human source, and the constant domain (if any) is from a human source.

[0061] The antibodies or fragments thereof of the present invention can 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 provide for the presence of any recited 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 "comprises" includes the more restrictive terms "consisting of" and "consisting essentially of". In one embodiment, the term "comprises" used throughout the application and particularly in the claims can be replaced with the term "consisting of".

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

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

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

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

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

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

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

[0070] vi) In the case of incorporating a heterologous amino acid sequence into CDRL3, between the amino acid residue LC res88 according to Kabat (in the light chain having the amino acid sequence SEQ ID NO: 66, which corresponds to res C90) 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 the following

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

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

[0074] iii) In the case of incorporating a heterologous amino acid sequence into CDRH3, according to Kabat, amino acid residue HC res106 (in the heavy chain having the amino acid sequence SEQ ID NO: 65, which corresponds to res G111);

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

[0076] v) In the case of incorporating a heterologous amino acid sequence into CDRL2, according to Kabat, amino acid residue LC res57 (in the light chain having the amino acid sequence SEQ ID NO: 66, which corresponds to res G59); and / or

[0077] vi) In the case of incorporating a heterologous amino acid sequence into CDRL3, according to Kabat, amino acid residue LC res98 (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 positions in the original immunoglobulin molecule before the incorporation 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 (the standard CDR boundary definitions are amino acid residues S25 and W36 for CDRH1; S49 and R67 for CDRH2; K98 and W108 for CDRH3; C22 and W37 for CDRL1; Y51 and G59 for CDRL2; C90 and F102 for CDRL3. Jarasch and Skerra, Proteins 2017 Jan; 85(1): 65 - 71).

[0079] The reference aa that is the nearest neighbor to the N - terminus of the inserted natriuretic peptide plus the amino acid segment present between said 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 contains Ntls. In a specific embodiment, the N - terminal sequence consists of Ntls plus the adjacent N - terminal reference aa.

[0080] The amino acid segment present between the last amino acid residue of the inserted natriuretic peptide and the reference aa that is the C-terminal nearest neighbor of the inserted natriuretic peptide, plus said reference aa, is herein referred to as the "C-terminal sequence". The C-terminal sequence contains 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 the human IgG antibody scaffold, or a sequence having at least 80% sequence identity thereto, particularly an amino acid sequence that is part of the fab domain scaffold of a human IgG antibody, or a sequence having at least 80% sequence identity thereto, more particularly any one of the sequences of SEQ ID NO 1, 2 or 4, or a sequence having at least 80% sequence identity to any one of SEQ ID NO 1, 2 or 4; any one of the sequences of SEQ ID NO 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 to any one of SEQ ID NO 6, 7, 9, 11, 13, 15, 16, 17, 19 or 21. Ntls may also comprise any combination of the amino acid sequences listed above.

[0082] In a specific such embodiment, Ntls comprises a GS linker sequence; a PN linker sequence; an amino acid sequence that is part of the human IgG antibody scaffold, or a sequence having at least 80% sequence identity thereto, particularly an amino acid sequence that is part of the fab domain scaffold of a human IgG antibody or a sequence having at least 80% sequence identity thereto, more particularly any one of the sequences of SEQ ID NO 1, 2 or 4, or a sequence having at least 80% sequence identity to any one of SEQ ID NO 1, 2 or 4; any one of the sequences of SEQ ID NO 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 to any one of SEQ ID NO 6, 7, 9, 11, 13, 15 or 21; or any combination thereof.

[0083] In a specific embodiment, Ctls comprise a GS linker sequence; a PN linker sequence; an amino acid sequence that is part of the human IgG antibody scaffold or a sequence having at least 80% sequence identity thereto, particularly an amino acid sequence that is part of the fab domain scaffold of a human IgG antibody or a sequence having at least 80% sequence identity thereto, more particularly any one of the sequences of SEQ ID NO: 1, 3 or 5, or a sequence having at least 80% sequence identity to any one of SEQ ID NO: 1, 3 or 5; any one of the sequences of SEQ ID NO: 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 to any one of SEQ ID NO: 6, 8, 10, 12, 14, 15, 17, 18, 19, 20 or 22. Ctls may also comprise any combination of the amino acid sequences listed above.

[0084] In a specific embodiment, Ctls comprise a GS linker sequence; a PN linker sequence; an amino acid sequence that is part of the human IgG antibody scaffold or a sequence having at least 80% sequence identity thereto, particularly an amino acid sequence that is part of the fab domain scaffold of a human IgG antibody or a sequence having at least 80% sequence identity thereto, more particularly any one of the sequences of SEQ ID NO: 1, 3 or 5, or a sequence having at least 80% sequence identity to any one of SEQ ID NO: 1, 3 or 5; any one of the sequences of SEQ ID NO: 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 to any one of SEQ ID NO: 6, 8, 10, 12, 14, 15, 20 or 22; or any combination thereof.

[0085] In a specific embodiment, the sequence identity between the sequences comprised in Ntls and / or Ctls and any one of SEQ ID NOs: 1 to 22 is 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%.

[0086] In the context of the present invention, the term "GS linker sequence" refers to a peptide linker mainly comprising 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 in total 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 residue that is not glycine or serine.

[0087] In the context of the present invention, the term "PN linker sequence" refers to a peptide linker mainly comprising 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 in total 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 residue that is 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 glutamate residues.

[0088] In a specific embodiment, a linker sequence contained within Ntls and / or Ctls and selected from the following comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid residues: GS linker sequence; PN linker sequence; human IgG antibody scaffold linker sequence; human IgG fab domain scaffold sequence; a sequence having at least 80% sequence identity with a human IgG antibody scaffold linker sequence, a human IgG fab domain scaffold sequence or any one of SEQ ID NOs: 1 to 5; a sequence having at least 60% sequence identity with 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 therewith, it may be particularly advantageous to use the sequence of the scaffold region adjacent to the CDR incorporating the heterologous amino acid sequence. For example, in the case where the heterologous amino acid sequence is incorporated into the CDRH2 domain, the Ntls and / or Ctls may comprise a linker containing an amino acid sequence that is part of the framework region FRH2 or FRH3. Similarly, in the case where the heterologous amino acid sequence is incorporated into the CDRL2 region, the Ntls and / or Ctls may comprise a linker containing an amino acid sequence that is part of the framework region FRH2 or FRH3.

[0090] In a specific embodiment, the Ntls consists of: 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 therewith, particularly an amino acid sequence that is part of the fab domain scaffold of a human IgG antibody or a sequence having at least 80% sequence identity therewith, more particularly any one of the sequences of SEQ ID NO 1, 2 or 4 or a sequence having at least 80% sequence identity with any one of SEQ ID NO 1, 2 or 4; any one of the sequences of SEQ ID NO 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 with any one of SEQ ID NO 6, 7, 9, 11, 13, 15, 16, 17, 19 or 21; or any combination thereof.

[0091] In a specific embodiment, the Ctls consists of: 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 therewith, particularly an amino acid sequence that is part of the fab domain scaffold of a human IgG antibody or a sequence having at least 80% sequence identity therewith, more particularly any one of the sequences of SEQ ID NO 1, 3 or 5 or a sequence having at least 80% sequence identity with any one of SEQ ID NO 1, 3 or 5; any one of the sequences of SEQ ID NO 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 with any one of SEQ ID NO 6, 8, 10, 12, 14, 15, 17, 18, 19, 20 or 22; or any combination thereof.

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

[0093] When the total length of the N-terminal and C-terminal flanking sequences is sufficient, the above-listed linker sequences have proven to be particularly advantageous for obtaining good natriuretic peptide activity. Without wishing to be bound by theory, it is believed that the above-listed linker peptides result in such a conformation / folding that it contributes to a systemic state that favors presenting the bioactive natriuretic peptide to its respective receptor with minimal steric hindrance.

[0094] In a specific embodiment, i) Ntls comprises a GS linker sequence; a PN linker sequence; the sequences of SEQ ID NOs: 2, 4, 9, 11, 13 or 15; a sequence having at least 60% sequence identity with SEQ ID NOs: 2, 4, 9, 11, 13 or 15; or any combination thereof, and ii) Ctls comprises a GS linker sequence; a PN linker sequence; the sequences of SEQ ID NOs: 3, 5, 12, 14, 15 or 20; a sequence having at least 60% sequence identity with SEQ ID NOs: 3, 5, 12, 14, 15 or 20; or any combination thereof. These linker sequences have proven to be particularly useful because they not only achieve high natriuretic peptide activity but also have good expression levels in recombinant expression and are not prone to protein cleavage upon expression (see Table 9).

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

[0096] In certain such embodiments, each of Ntls and Ctls comprises a GS linker sequence; each of Ntls and Ctls comprises a PN linker sequence; each of Ntls and Ctls comprises 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 the fab domain scaffold of a human IgG antibody or a sequence having at least 80% sequence identity thereto, more specifically Ntls comprises any one of the sequences of SEQ ID NO: 1, 2 or 4 or a sequence having at least 80% sequence identity to any one of SEQ ID NO: 1, 2 or 4, and Ctls comprises any one of the sequences of SEQ ID NO: 1, 3 or 5 or a sequence having at least 80% sequence identity thereto; each of Ntls and Ctls comprises the sequence SEQ ID NO: 6 or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence SEQ ID NO: 7 or a sequence having at least 60% sequence identity thereto, and Ctls comprises the sequence SEQ ID NO: 8 or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence SEQ ID NO: 9 or a sequence having at least 60% sequence identity thereto, and Ctls comprises the sequence SEQ ID NO: 10 having at least 60% sequence identity thereto; Ntls comprises the sequence SEQ ID NO: 11 or a sequence having at least 60% sequence identity thereto, and Ctls comprises the sequence SEQ ID NO: 12 or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence SEQ ID NO: 13 or a sequence having at least 60% sequence identity thereto, and Ctls comprises the sequence SEQ ID NO: 14 or a sequence having at least 60% sequence identity thereto; each of Ntls and Ctls comprises the sequence SEQ ID NO: 15 or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence SEQ 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; or Ntls comprises the sequence SEQ ID NO: 21 or a sequence having at least 60% sequence identity thereto, and Ctls comprises the sequence 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 SEQ ID NO 2 or a sequence having at least 80% sequence identity thereto, and Ctls comprises the sequence SEQ ID NO 3 or a sequence having at least 80% sequence identity thereto; Ntls comprises the sequence SEQ ID NO 4 or a sequence having at least 80% sequence identity thereto, and Ctls comprises the sequence SEQ ID NO 5 or a sequence having at least 80% sequence identity thereto; Ntls comprises the sequence SEQ ID NO 11 or a sequence having at least 60% sequence identity thereto, and Ctls comprises the sequence SEQ ID NO 12 or a sequence having at least 60% sequence identity thereto; Ntls comprises the sequence SEQ ID NO 13 or a sequence having at least 60% sequence identity thereto, and Ctls comprises the sequence SEQ ID NO 14 or a sequence having at least 60% sequence identity thereto; Ntls and Ctls each comprise the sequence SEQ ID NO 15 or a sequence having at least 60% sequence identity thereto; or Ntls comprises the sequence SEQ ID NO 9 or a sequence having at least 60% sequence identity thereto, 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 a specific embodiment, Ntls further comprises an anchoring element A1 at its C-terminal end and / or Ctls further comprises an anchoring element A2 at its N-terminal end, wherein A1 and A2 mainly comprise glycine and serine residues. In a specific embodiment, 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 a specific embodiment, 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 that are not 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 the human IgG antibody scaffold or a sequence having at least 80% sequence identity thereto, particularly an amino acid sequence that is part of the fab domain scaffold of a human IgG antibody or a sequence having at least 80% sequence identity thereto, more particularly any one of the sequences of SEQ ID NO 1, 2 or 4 or a sequence having at least 80% sequence identity to any one of SEQ ID NO 1, 2 or 4; any one of the sequences of SEQ ID NO 6, 7, 9, 11, 13, 15, 16, 17, 19 or 21; a sequence having at least 60% sequence identity to any one of the sequences of SEQ ID NO 6, 7, 9, 11, 13, 15, 16, 17, 19 or 21; 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 the human IgG antibody scaffold or a sequence having at least 80% sequence identity thereto, particularly an amino acid sequence that is part of the fab domain scaffold of a human IgG antibody or a sequence having at least 80% sequence identity thereto, more particularly any one of the sequences of SEQ ID NO 1, 3 or 5 or a sequence having at least 80% sequence identity to any one of SEQ ID NO1, 3 or 5; any one of the sequences of SEQ ID NO 6, 8, 10, 12, 14, 15, 17, 18, 19, 20 or 22; a sequence having at least 60% sequence identity to any one of the sequences of SEQ ID NO 6, 8, 10, 12, 14, 15, 17, 18, 19, 20 or 22; or any combination thereof.

[0101] In a specific embodiment, Ntls and / or Ctls together contain at least 3, 4, 5, 6, 7, 8, 9 or 10 amino acid residues. Ntls and / or Ctls can, for example, together contain 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.

[0102] In a specific embodiment, the amino acid segment present in the following contains any one of the sequences of SEQ ID NO 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 NO 26 to 38

[0103] i) When incorporating the heterologous amino acid sequence into CDRH1, between the amino acid residue HCres25 according to Kabat and the first amino acid residue of the natriuretic peptide;

[0104] ii) When incorporating the heterologous amino acid sequence into CDRH2, between the amino acid residue HCres51 according to Kabat and the first amino acid residue of the natriuretic peptide;

[0105] iii) When incorporating the heterologous amino acid sequence into CDRH3, between the amino acid residue HCres92 according to Kabat and the first amino acid residue of the natriuretic peptide;

[0106] iv) When incorporating the heterologous amino acid sequence into CDRL1, between the amino acid residue LCres26 according to Kabat and the first amino acid residue of the natriuretic peptide;

[0107] v) When incorporating the heterologous amino acid sequence into CDRL2, between the amino acid residue LCres49 according to Kabat and the first amino acid residue of the natriuretic peptide; and / or

[0108] vi) When incorporating the heterologous amino acid sequence into CDRL3, between the amino acid residue LCres88 according to Kabat and the first amino acid residue of the natriuretic peptide.

[0109] In a specific embodiment, the amino acid segment present between the last amino acid residue of the natriuretic peptide and the following contains 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 with any one of SEQ ID NOs: 39 to 51

[0110] i) When incorporating the heterologous amino acid sequence into CDRH1, according to the amino acid residue HCres35a of Kabat;

[0111] ii) When incorporating the heterologous amino acid sequence into CDRH2, according to the amino acid residue HCres57 of Kabat;

[0112] iii) When incorporating the heterologous amino acid sequence into CDRH3, according to the amino acid residue HCres106 of Kabat;

[0113] iv) When incorporating the heterologous amino acid sequence into CDRL1, according to the amino acid residue LCres 32 of Kabat;

[0114] v) In the case of incorporating the heterologous amino acid sequence into CDRL2, the amino acid residue LCres57 according to Kabat; and / or

[0115] vi) In the case of incorporating the heterologous amino acid sequence into CDRL3, the amino acid residue LCres98 according to Kabat.

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

[0117] In a specific embodiment, the amino acid segment present in the following contains 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 with any one of SEQ ID NOs: 52 to 64

[0118] i) Between the amino acid residue HCres25 according to Kabat and the amino acid residue HC res35a according to Kabat in the case of incorporating the heterologous amino acid sequence into CDRH1;

[0119] ii) Between the amino acid residue HCres51 according to Kabat and the amino acid residue HC res57 according to Kabat in the case of incorporating the heterologous amino acid sequence into CDRH2;

[0120] iii) Between the amino acid residue HCres92 according to Kabat and the amino acid residue HC res106 according to Kabat in the case of incorporating the heterologous amino acid sequence into CDRH3;

[0121] iv) Between the amino acid residue LCres26 according to Kabat and the amino acid residue LC res 32 according to Kabat in the case of incorporating the heterologous amino acid sequence into CDRL1;

[0122] v) Between the amino acid residue LCres49 according to Kabat and the amino acid residue LC res57 according to Kabat in the case of incorporating the heterologous amino acid sequence into CDRL2; and / or

[0123] vi) Between the amino acid residue LCres88 according to Kabat and the amino acid residue LC res98 according to Kabat in the case of incorporating the heterologous amino acid sequence into CDRL3.

[0124] In a specific embodiment, the antibody or fragment thereof comprises at least two natriuretic peptides. In a specific embodiment, both of the two natriuretic peptides are included in a heterologous amino acid sequence that further comprises Ntls and Ctls and are incorporated into the CDR regions of the antibody or fragment thereof 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 such embodiment, the antibody or fragment thereof comprises at least two different natriuretic peptides that are incorporated into at least two CDR regions.

[0125] Due to the dimeric structure of the antibody molecule, insertion of a nucleic acid sequence encoding a heterologous amino acid sequence (Ntls-natriuretic peptide-Ctls) into the nucleic acid encoding the light or heavy chain of an immunoglobulin molecule typically results in an antibody protein carrying two natriuretic peptides located in the corresponding CDR regions of two identical light chains or two identical heavy chains. However, it is also contemplated to insert two nucleic acids encoding natriuretic peptides into two different CDR-encoding regions of the nucleic acid sequence encoding the light and / or heavy chain, thereby generating an antibody molecule having four natriuretic peptides located in two corresponding CDR pairs of the dimeric antibody. Also included are dimeric immunoglobulin molecules with different light and / or heavy chains, 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 a specific embodiment, 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 a specific embodiment, the antibody or fragment thereof comprises an ANP and a BNP molecule; an ANP and a CNP molecule; or a BNP and a CNP molecule.

[0127] In a specific embodiment, the natriuretic peptide in at least one heterologous amino acid sequence incorporated into at least one CDR region of the antibody or its fragment is ANP, and the antibody or its fragment comprises at least one other natriuretic peptide. In a specific embodiment, the at least one other natriuretic peptide is also included in the heterologous amino acid sequence, which further comprises Ntls and Ctls and is incorporated into the CDR region of the antibody or its fragment. In a specific embodiment, ANP 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, ANP 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, and more specifically from BNP and CNP.

[0128] A "blank" antibody molecule consisting of two heavy chains having the sequence SEQ ID NO: 65 and two light chains having the sequence 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 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 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 antibody or parental antibody, and a heterologous amino acid sequence comprising a natriuretic peptide is incorporated therein. In a specific such embodiment, the heterologous amino acid sequence comprising a natriuretic peptide is incorporated into the CDR region of one or two heavy chains of such an antibody molecule.

[0129] In a specific such embodiment, a heavy chain comprising at least one heterologous amino acid sequence incorporated into at least one of its CDR regions has a 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 a specific embodiment, the antibody of the present invention consists of two identical heavy chains and two identical light chains, the heavy chain having a 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 the light chain having the sequence 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 regions of one or both light chains of such antibody molecules. In specific such embodiments, a light chain comprising at least one heterologous amino acid sequence incorporated into at least one of its CDR regions has the sequence of either SEQ ID NO: 80 or 81 or a sequence having at least 80%, at least 85%, at least 90% or at least 95% sequence identity to either SEQ ID NO: 80 or 81. In a specific embodiment, an antibody of the invention consists of two identical light chains and two identical heavy chains, wherein the light chain has the sequence of either SEQ ID NO: 80 or 81 or a sequence having at least 80%, at least 85%, at least 90% or at least 95% sequence identity to either SEQ ID NO: 80 or 81, and the heavy chain has the sequence 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 compositions 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 invention include naturally occurring purified products, products of chemical synthesis steps, and products produced by recombinant techniques. Depending on their origin, the antibodies or fragments thereof of the invention can be glycosylated or non-glycosylated.

[0136] For example, standard recombinant DNA methods can be used to prepare and / or obtain nucleic acids encoding heavy and light chains, incorporate these nucleic acids into an expression vector, and introduce the vector into a host cell for recombinant expression (see, e.g., Sambrook, Fritsch and Maniatis (eds.), Molecular Cloning; A Laboratory Manual, Second Edition, Cold Spring Harbor, N.Y., (1989); Ausubel, F.M. 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. 4,816,397 to Boss et al.).

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

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

[0139] Useful expression vectors for bacterial use can be constructed by inserting one or more nucleic acid sequences of the invention, appropriate translation initiation and termination signals, together with a functional promoter, into an operable reading phase. Bacterial expression vectors generally include one or more phenotypic selection markers and an origin of replication to ensure maintenance of the vector and, if desired, provide amplification in a bacterial host. Bacterial expression vectors may contain elements derived from commercially available plasmids, such as the well-known cloning vector pBR322 (ATCC 37017). Depending on the intended use of the antibody or fragment thereof to be expressed, many bacterial expression vectors can be advantageously selected. For example, if large amounts of such antibodies are needed, a vector that mediates high-level expression of an antibody fusion protein that is easily purified may be required.

[0140] A recombinant nucleic acid construct intended for antibody expression in eukaryotic host cells may comprise regulatory sequences capable of regulating the expression of an open reading frame in eukaryotic cells, preferably a promoter and a polyadenylation signal. The promoter and polyadenylation signal are preferably selected to be functional in the particular cell type intended for antibody expression. Examples of suitable promoters include, but are not limited to, promoters from cytomegalovirus (CMV) (such as the strong CMV immediate early promoter), simian virus 40 (SV40), mouse mammary tumor virus (MMTV), human immunodeficiency virus (HIV) (such as the HIV long terminal repeat (LTR) promoter), Moloney virus, Epstein - Barr virus (EBV), adenovirus (such as the adenovirus major late promoter (ADMLP)), polyoma, and Rous sarcoma virus (RSV); the synthetic CAG promoter consisting of the CMV early enhancer element, the promoter of the chicken β - actin gene, the first exon and the first intron, 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 a specific embodiment, the eukaryotic expression cassette comprises the 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 can 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 their sequences, see US 5,168,062 by Stinski, US 4,510,245 by Bell et al., and US 4,968,615 by Schaffner et al.

[0143] Regulatory sequences and codons are generally species - dependent, and thus, to maximize protein production, regulatory sequences and codons effective in the species / cell type intended for antibody expression are preferably selected. Those skilled in the art can produce recombinant DNA molecules 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 auxotrophies (such as glutamine synthase) on the host cells into which the vector has been introduced. For example, the dihydrofolate reductase (DHFR) gene confers resistance to methotrexate, the neo 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 zeocin, and the Escherichia coli hygromycin resistance gene (hyg or hph) confers resistance to hygromycin. Selectable markers such as DHFR or glutamine synthase can also be used in combination with MTX and MSX for amplification techniques.

[0145] In some embodiments, nucleic acid sequences encoding the heavy and light chains are inserted into separate vectors. In other embodiments, nucleic acid sequences encoding the heavy and light chains are inserted into the same vector. Additionally, nucleic acid sequences encoding the variable regions of the heavy and / or light chains can be converted to, for example, nucleic acid sequences encoding full-length antibody chains, Fab fragments, or scFvs. DNA fragments encoding VL or VH can be operably linked (such that the amino acid sequences encoded by the two DNA fragments are 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 fragment encoding a flexible linker such that the VH and VL sequences can be expressed as a continuous single-chain protein and the VL and VH regions are joined by the 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). Sequences of the human heavy and light chain constant regions are known in the art (see, e.g., Kabat, E.A., el al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242), and DNA fragments containing these regions can be obtained by standard PCR amplification.

[0146] In a specific embodiment, the nucleic acid sequence encoding the heavy chain incorporating a heterologous amino acid sequence comprising a natriuretic peptide comprises the sequence of either SEQ ID NO 82 or 83, or a sequence having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity to either SEQ ID NO 82 or 83. In a specific such embodiment, 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 invention relates to a host cell comprising the nucleic acid or nucleic acid mixture of the invention. In the context of the present invention, the terms "host cell", "host cell line" and "host cell culture" are used interchangeably and refer to a cell into which exogenous nucleic acid has been introduced, including progeny of such cells. Host cells include "transformants", "transformed cells", "transfectants", "transfected cells" and "transduced cells", which include the primary transformed / transfected / transduced cells and progeny derived therefrom, regardless of the number of passages. The nucleic acid content of the progeny may not be identical to that of the parental cell, and the exogenous nucleic acid contained may contain mutations. Mutant progeny having the same function or biological activity as that screened or selected in the originally transformed cell are included herein.

[0148] Transfection of the expression vector into the host cell can be carried out 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 and eukaryotic cells. Examples of prokaryotic host cells are, for example, bacteria including, but not limited to, Escherichia coli, Bacillus subtilis, Salmonella typhimurium, and various species in the genera 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 as 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 selectable marker, such as as described in R.J. Kaufman and P.A. Sharp (1982) Mol. Biol. 159:601-621; and other knockout cells as exemplified 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 in the expression system can also be transient or semi-stable, such as in 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 cells of the present invention. Specifically, the host cells of the present invention are cultured under conditions suitable for the expression of the antibody or a fragment thereof.

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

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

[0155] In a specific embodiment, the method of the present invention further includes the step of purifying the recovered antibody or fragment thereof. 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 instrument), more specifically to produce an antibody homogeneity of at least about 92.5%, 95%, 98% or 99%; or, the antibody is purified (2) to an extent sufficient to obtain at least 15 residues of the N-terminal or internal amino acid sequence, or (3) to homogeneity by SDS-PAGE using Coomassie blue or preferably silver staining under reducing or non-reducing conditions.

[0156] The antibody or fragment 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, N.Y., (1997-2001), e.g., Chapters 1, 4, 6, 8, 9, 10).

[0157] In a fifth aspect, the present invention relates to a composition comprising the antibody or fragment thereof of the present invention. Specifically, the composition of the present invention is a pharmaceutical composition suitable for use in a method of treatment, wherein the amount of the antibody or fragment thereof of the present invention contained therein is 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 together with the active ingredient of a drug. Suitable excipients include anti-adhesives, binders, coatings, disintegrants, flavorings, pigments, lubricants, glidants, adsorbents, preservatives, and sweetening agents. 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" means that a molecular entity and other components of a pharmaceutical composition are physiologically tolerable and generally do not produce adverse reactions when administered to a mammal (e.g., a human). The term "pharmaceutically acceptable" may also refer to being approved by a regulatory agency of the federal or state government or listed in the United States Pharmacopeia or other generally recognized pharmacopeias for mammals (more specifically humans).

[0159] The composition of the present invention may further comprise one or more other therapeutic 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] Pharmaceutically acceptable excipients known in the art can be used to formulate a pharmaceutical composition for oral administration in a dosage suitable for oral administration. Such excipients enable the pharmaceutical composition to be formulated into tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc. for ingestion by a patient.

[0162] A pharmaceutical product for oral use can be obtained by combining the active compound with a solid excipient, optionally grinding the resulting mixture, and processing the particulate mixture with the addition of suitable auxiliaries (if necessary) to obtain tablets or dragee cores. Suitable excipients are carbohydrate or protein fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; starches from corn, wheat, rice, potato, or other plants; celluloses such as methylcellulose, hydroxypropylmethylcellulose, or sodium carboxymethylcellulose; and gums, including gum arabic and tragacanth; and proteins such as gelatin and collagen. If necessary, disintegrants or solubilizers such as cross-linked polyvinylpyrrolidone, agar, alginic acid, or its salts such as sodium alginate can be added.

[0163] The dragee cores can be provided with suitable coatings (such as concentrated sugar solutions), which may also contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquear solution, and suitable organic solvents or solvent mixtures. Dyes or pigments can be added to the tablet or dragee coatings for product identification or to characterize the amount of the active compound, i.e., the dose.

[0164] Oral dosage forms of the pharmaceutical compositions include push-fit capsules made of gelatin as well as soft-sealed capsules made of gelatin and a coating such as glycerol or sorbitol. The push-fit capsules may contain the active ingredient admixed with fillers or binders such as lactose or starch, lubricants such as talc or magnesium stearate, and optionally stabilizers. In the soft gelatin capsules, the active compounds may be dissolved or suspended in a suitable liquid such as a fatty oil, liquid paraffin or liquid polyethylene glycol with or without stabilizers.

[0165] Parenteral dosage forms of the pharmaceutical compositions include aqueous solutions of the therapeutic active agent. For injection, the pharmaceutical compositions of the invention may be formulated in an aqueous solution, preferably in a physiologically compatible buffer such as Hank's solution, Ringer's solution or physiological saline buffer. The aqueous injection suspensions may contain substances that increase the viscosity of the suspension such as sodium carboxymethyl cellulose, sorbitol or dextran. Additionally, suspensions of the active agent may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or carriers include fatty oils such as sesame oil, synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the therapeutic active agent to allow for the preparation of highly concentrated solutions.

[0166] For topical or nasal administration, permeants suitable for the particular barrier to be permeated are used in the dosage form. Such permeants are generally known in the art.

[0167] The pharmaceutical compositions of the invention may be manufactured in a manner known in the art, e.g., by conventional mixing, dissolving, granulating, sugar coating, suspending, emulsifying, encapsulating, entrapping or lyophilizing processes.

[0168] The pharmaceutical compositions may be provided in the form of salts and formed with acids including, but not limited to, hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, etc. The salts tend to be more soluble in aqueous or other protonic solvents than are the corresponding free base forms. In other cases, the preferred dosage form 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] Other details of the dosage forms and administration techniques can be found in the latest edition of Remington's Pharmaceutical Sciences (Ed. Maack Publishing Co, Easton, Pa.).

[0170] After preparing the pharmaceutical composition containing the antibody or fragment thereof of the invention, it may be placed in a suitable container and labeled for the specified disorder for which it is to be used in treatment. Such labeling for administration of the antibody or fragment of the invention will include the dosage amount, frequency and method.

[0171] The present invention also provides a pack and a kit comprising one or more containers containing one or more ingredients of the foregoing compositions of the present invention. Accompanying such containers is a notice 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 the use of an antibody or a fragment thereof of the present invention or a composition of the present invention in a method of treatment.

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

[0174] In the context of the present invention, the term "therapeutically effective amount" is defined as the amount of an antibody or a fragment thereof of the present invention sufficient to prevent or alleviate the disease symptoms of any disorder and disease mentioned herein, as a single dose or according to a multi-dose regimen, alone or in combination with other medicaments. In a specific embodiment, the "therapeutically effective amount" is toxicologically tolerable. Determination of an effective dose is entirely 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, gender, and disease state, the time, frequency, and route of administration, the drug combination, and the nature of the disorder being treated. Usual doses of antibodies vary from 0.1 micrograms to 100,000 micrograms, up to a total dose of about 10 g, depending on the route of administration.

[0175] For example, general guidance on methods for its determination can be found in the publications of the International Conference on Harmonization and in Chapters 27 and 28 of REMINGTON′S PHARMACEUTICAL SCIENCES, pages 484 - 528 (18th ed., Alfonso R. Gennaro, Ed., Easton, Pa.: Mack Pub. Co., 1990). More specifically, determination of 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 foregoing references. In general, the therapeutic efficacy and toxicity of a therapeutic agent can be determined in cell culture assays or animal models, such as the ED50 (the dose therapeutically effective in 50% of the population) and the LD50 (the dose lethal to 50% of the population). The ratio of the ED50 to the LD50 dose is the therapeutic index.

[0176] The antibodies or fragments thereof of the present invention are suitable for the treatment and / or prevention of 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 for the treatment and / or prevention of any one or any combination of these disorders and diseases.

[0177] Accordingly, the antibodies or fragments thereof of the present invention can be used in medicaments for the treatment and / or prevention of cardiovascular conditions such as arterial and pulmonary hypertension, refractory and resistant hypertension, acute and chronic heart failure, coronary heart disease, bronchiolitis obliterans syndrome (BOS), tumor-related and oncology diseases, graft-versus-host disease, sickle cell disease, stable and unstable angina, peripheral and cardiac vascular disorders, arrhythmias, atrial and ventricular arrhythmias and impaired conduction (such as atrioventricular block degrees I-III (AB block I-III)), supraventricular tachyarrhythmias, atrial fibrillation, atrial flutter, ventricular fibrillation, ventricular flutter, ventricular tachyarrhythmias, torsades de pointes, atrial and ventricular premature contractions, AV junctional premature contractions, sick sinus syndrome, syncope, atrioventricular nodal reentrant tachycardia, Wolff-Parkinson-White syndrome, acute coronary syndrome (ACS), autoimmune heart diseases (pericarditis, endocarditis, valvulitis, aortitis, cardiomyopathy), shock (such as cardiogenic shock, septic shock and anaphylactic shock), aneurysms, boxer cardiomyopathy (ventricular premature contractions (PVC)); can be used in medicaments 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 conditions, coronary and peripheral artery spasm, edema formation (such as pulmonary edema, cerebral edema, renal edema or edema due to heart failure), peripheral circulatory disorders, reperfusion injury, arterial and venous thrombosis, microalbuminuria, myocardial dysfunction, endothelial dysfunction, prevention of restenosis after, for example, thrombolytic therapy, percutaneous transluminal angioplasty (PTA), percutaneous transluminal coronary angioplasty (PTCA), heart transplantation and bypass surgery, and microvascular and macrovascular injury (vasculitis), elevated fibrinogen and low density lipoprotein (LDL) levels, elevated plasminogen activator inhibitor 1 (PAI-1) concentration; and can be used in medicaments 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, congestive heart failure, ischemic cardiomyopathy, dilated cardiomyopathy, hypertrophic cardiomyopathy, idiopathic cardiomyopathy, congenital heart defects, heart failure related to cardiac valve defects, mitral stenosis, mitral insufficiency, aortic stenosis, aortic insufficiency, tricuspid stenosis, tricuspid insufficiency, pulmonary valve stenosis, pulmonary valve insufficiency, combined cardiac 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 the acute exacerbation phase of existing chronic heart failure (worsening heart failure).

[0179] In addition, the antibodies or fragments thereof of the present invention can also be used for the treatment and / or prevention of arteriosclerosis, peripheral artery 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 for the treatment and / or prevention of primary and secondary Raynaud's phenomenon, impaired microcirculation, claudication, peripheral and autonomic neuropathy, diabetic microangiopathy, diabetic retinopathy, diabetic foot ulcers, gangrene, CREST syndrome, erythromelalgia, onychomycosis, rheumatism, and for promoting 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 enlargement (BPE), bladder outlet obstruction (BOO), lower urinary tract syndrome (LUTS, including feline urological syndrome (FUS)), genitourinary disorders including overactive bladder (OAB) and interstitial cystitis (IC), urinary incontinence (UI) (such as mixed urinary incontinence, urge urinary incontinence, stress urinary incontinence, or overflow urinary incontinence (MUI, UUI, SUI, OUI)), pelvic pain, and benign and malignant disorders of male and female genitourinary organs.

[0182] The antibodies or fragments thereof of the present invention are also suitable for the treatment and / or prevention of kidney 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 underlying or related kidney disorders such as renal hypoperfusion, hypotension during dialysis, obstructive uropathy, glomerulopathy, glomerulonephritis, acute glomerulonephritis, glomerulosclerosis, tubulointerstitial disease, nephrotic disorders (such as primary and congenital kidney diseases), nephritis, immune kidney disorders (such as kidney transplant rejection and immune complex-induced kidney disorders, toxin-induced nephropathy, contrast-induced nephropathy, diabetic and non-diabetic nephropathy, pyelonephritis, renal cysts, renal sclerosis, hypertensive renal sclerosis and nephrotic syndrome, which may be diagnostically characterized by, for example, abnormally reduced creatinine and / or water excretion, abnormally elevated urea, nitrogen, potassium and / or creatinine blood concentrations, altered renal enzyme (such as glutamine synthetase) activity, altered urine osmolality or urine volume, elevated microalbuminuria, macroalbuminuria, glomerular and arteriolar lesions, tubule dilation, 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 disorders (such as hyperkalemia, hyponatremia) and bone and carbohydrate metabolism disorders.

[0183] In addition, the antibodies or fragments thereof of the present invention are also suitable for the treatment and / or prevention of asthma disorders, pulmonary arterial hypertension (PAH) and other forms of pulmonary hypertension (PH) including left heart disease, HIV, sickle cell anemia, chronic thromboembolic pulmonary hypertension (CTEPH), sarcoidosis, COPD- or pulmonary fibrosis-related pulmonary hypertension, chronic obstructive pulmonary disease (COPD), acute respiratory distress syndrome (ARDS), acute lung injury (ALI), alpha-1-antitrypsin deficiency (AATD), pulmonary fibrosis, emphysema (such as 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 disorders of the NO / cGMP system. It is particularly suitable for improving perception, attention, learning or memory after cognitive deficits, such as those deficits associated with, in particular, the following conditions / diseases / syndromes: such as mild cognitive deficits, age-related learning and memory deficits, age-related memory loss, vascular dementia, traumatic brain injury, stroke, dementia occurring after stroke (post-stroke dementia), post-traumatic brain injury, general attention disorders, attention disorders in children with learning and memory problems, Alzheimer's disease, dementia with Lewy bodies, frontotemporal dementia (including Pick's syndrome), Parkinson's disease, progressive supranuclear palsy, corticobasal ganglionic degeneration dementia, amyotrophic lateral sclerosis (ALS), Huntington's disease, demyelination, multiple sclerosis, thalamic degeneration, Creutzfeld-Jacob dementia, HIV dementia, schizophrenic dementia or Korsakoff psychosis. It is also suitable for treating and / or preventing central nervous system disorders, such as anxiety, tension and depressive states, CNS-related sexual dysfunction and sleep disorders, and for controlling pathological disorders of 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 thus effective agents for controlling migraine. It is also suitable for preventing and controlling the sequelae of cerebral infarction (stroke), such as stroke, cerebral ischemia and traumatic brain injury. It can also be used to control pain and tinnitus conditions.

[0186] In addition, the antibodies or fragments of the present invention have an anti-inflammatory effect and can thus be used as anti-inflammatory agents for treating and / or preventing 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 for treating and / or preventing autoimmune diseases.

[0188] The antibodies or fragments thereof are also suitable for treating and / or preventing fibrotic conditions of visceral organs such as the lung, heart, kidney, reproductive system, bone marrow, especially the liver, as well as skin 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 damage caused by diabetes, uterine fibroids, endometriosis, myelofibrosis and similar fibrotic disorders, scleroderma, morphea, keloids, hypertrophic scar formation (also after surgical procedures), moles, diabetic retinopathy, proliferative vitreoretinopathy and connective tissue disorders (such as sarcoidosis).

[0189] The antibody or fragment thereof of the present invention is also applicable to controlling postoperative scar formation, such as that caused by glaucoma surgery.

[0190] The antibody or fragment thereof of the present invention can also be used cosmetically for aged and keratinized skin.

[0191] In addition, the antibody or fragment thereof of the present invention is applicable to the treatment and / or prevention of hepatitis, tumors, osteoporosis, glaucoma, and gastroparesis.

[0192] In addition, the antibody or fragment thereof of the present invention is applicable to the treatment and / or prevention of 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, elevated 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 antibody or fragment thereof of the present invention is applicable to the treatment of bone and cartilage disorders, such as bone and cartilage diseases responsive to natriuretic peptides, arthritis, degenerative diseases of cartilage tissue, osteoarthritis, cartilage degeneration, fractures, skeletal dysplasia, achondroplasia, osteoporosis, osteogenesis imperfecta, Paget's disease of bone (PDB), metabolic bone diseases, age-related bone diseases, osteomyelitis, osteonecrosis, rickets, osteomalacia, growth plate injuries and diseases, defects related to joint and bone replacement, Marfan syndrome, sports injuries, muscular dystrophy, Duchenne muscular dystrophy.

[0194] Therefore, in another aspect, the present invention relates to the use of the antibody or fragment thereof of the present invention for the treatment and / or prevention of disorders, particularly the above-mentioned disorders.

[0195] In a specific embodiment, the antibody or fragment thereof of the present invention can be used in methods for the treatment and / or prevention of 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 the antibody or fragment thereof of the present invention for the production of a medicament for the treatment and / or prevention of disorders, particularly the above-mentioned disorders.

[0197] In specific embodiments, the present invention relates to the use of the antibodies or fragments thereof of the present invention for the production of a medicament for treating and / or preventing heart failure, angina pectoris, hypertension, pulmonary hypertension, ischemia, vascular disorders, renal insufficiency, thromboembolic disorders, fibrotic disorders, dementia, arteriosclerosis, skeletal and bone disorders, ocular disorders, dwarfism, achondroplasia and erectile dysfunction.

[0198] In another aspect, the present invention relates to a method of treating and / or preventing a disorder, particularly the above-mentioned disorders, using an effective amount of at least one antibody or fragment thereof of the present invention.

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

[0200] The antibody or fragment thereof of the present invention can be administered as the sole agent or in combination with one or more additional therapeutic agents. In some cases, the antibody itself may be modified. For example, the antibody or fragment thereof can be conjugated to a chemical entity, such as to further enhance efficacy, stability and / or half-life. Specifically, the antibody or fragment thereof of the present invention can be pegylated and / or HESylated (hydroxyethyl starchylated).

[0201] Thus, in specific embodiments, the antibody or fragment thereof of the present invention is used in combination with at least one additional therapeutic agent in a treatment method, particularly 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 such a combination may be a fixed combination, a non-fixed combination or a component kit.

[0204] In the context of the present invention, the term "fixed combination" is used in a manner known to those skilled in the art and is defined as a combination in which, for example, a first active ingredient (such as one or more antibodies or fragments thereof of the present invention), and another active ingredient are present together in a unit dose or a single entity, such as a single-dose formulation. An example of a "fixed combination" is a pharmaceutical composition in which the first active ingredient and another active ingredient are present as a mixture for simultaneous administration, for example, in a formulation. Another example of a "fixed combination" is a pharmaceutical combination in which the first active ingredient and another active ingredient are present in a single unit rather than as a mixture. Thus, the present invention provides such pharmaceutical compositions that comprise at least one antibody or fragment thereof and at least one additional therapeutic agent, particularly 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 components" are used in a manner known to those skilled in the art and are defined as a combination in which the first active ingredient and another active ingredient are present in more than one unit, such as in separate dosage articles. An example of a non-fixed combination or a kit of components is a combination in which the first active ingredient and another active ingredient are present separately.

[0206] The components of a non-fixed combination or a kit of components can be administered separately, sequentially, simultaneously, concurrently, or in a chronologically staggered manner.

[0207] The antibody or fragment thereof of the present invention can be administered simultaneously with, before, or after the said other therapeutic active agent. In the context of the present invention, the term "simultaneously" means that the antibody or fragment thereof of the present invention and at least one other therapeutic active agent are administered on the same day, more particularly within 12 hours, and 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 active 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 active 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 reagents in the same treatment group for an indication, such as reagents for treating and / or preventing diseases and disorders related to the following: hypertension, heart failure, pulmonary hypertension, COPD, asthma, cystic fibrosis, achondroplasia, hyperphosphatemia, chronic kidney disease (CKD), soft tissue calcification, CKD-related calcification, non-CKD-related calcification, medial calcification including Monckeberg's medial sclerosis, atherosclerosis, intimal calcification, CKD-related cardiac hypertrophy, CKD-related renal osteodystrophy, osteoporosis, postmenopausal osteoporosis, type II diabetes, chronic kidney disease, aging, hypophosphaturia, hyperparathyroidism, vitamin D disorders, vitamin K deficiency, vitamin K antagonist anticoagulants, Kawasaki disease, ACDC (arterial calcification due to CD73 deficiency), GACI (generalized arterial calcification in infancy), IBGC (idiopathic basal ganglia calcification), PXE (pseudoxanthoma elasticum), rheumatoid arthritis, Singleton-Merten syndrome, β-thalassemia, calciphylaxis, heterotopic ossification, premature placental calcification, uterine calcification, calcified uterine fibroids, Fahr's disease, aortic valve microcalcification and calcification.

[0210] Preferred examples of other suitable 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, molsidomine or SIN-1, and inhaled NO;

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

[0213] - Antithrombotic reagents, such as and preferably platelet aggregation inhibitors, anticoagulants or fibrinolytic substances;

[0214] - Hypotensive active ingredients, such as and preferably calcium antagonists, angiotensin AII antagonists, ACE inhibitors, NEP inhibitors, vasopeptidase inhibitors, endothelin antagonists, renin inhibitors, α-blockers, β-blockers, mineralocorticoid receptor antagonists, rho kinase inhibitors and diuretics;

[0215] - Antiarrhythmic agents, such as and preferably sodium channel blockers, β-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] - Inotropic agents, such as cardiac glycosides (Dogoxin), β-adrenergic and dopaminergic agonists, such as isoproterenol, epinephrine, norepinephrine, dopamine, or dobutamine;

[0217] - Vasopressin receptor antagonists, such as and preferably conivaptan, tolvaptan, lixivaptan, mozavaptan, satavaptan, SR-121463, RWJ 676070, or BAY 86-8050, and the compounds described in WO 2010 / 105770, WO2011 / 104322, and WO 2016 / 071212;

[0218] - Active ingredients that alter lipid metabolism, such as and preferably thyroid receptor agonists, PCSK9 inhibitors, cholesterol synthesis inhibitors, such as and preferably HMG-CoA reductase inhibitors or squalene synthesis inhibitors, ACAT inhibitors, CETP inhibitors, MTP inhibitors, PPAR-α, PPAR-γ, and / or PPAR-δ agonists, cholesterol absorption inhibitors, lipase inhibitors, polymeric bile acid adsorbents, bile acid reabsorption inhibitors, and lipoprotein(a) antagonists.

[0219] - Anti-inflammatory agents, such as and preferably glucocorticoids, such as and preferably prednisone, prednisolone, methylprednisolone, triamcinolone, dexamethasone, beclomethasone, betamethasone, flunisolide, budesonide, or fluticasone, and non-steroidal anti-inflammatory agents (NSAIDs), such as 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 that 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, such as and preferably matrix metalloproteinase (MMP) inhibitors, such as and preferably chymotrypsin, stromelysine, collagenase, gelatinase, and aggrecanase (preferably MMP-1, MMP-3, MMP-8, MMP-9, MMP-10, MMP-11, and MMP-13), as well as inhibitors of metalloelastase (MMP-12) and neutrophil elastase (HNE), such as sivelestat or DX-890;

[0222] - Agents that block the binding of serotonin to its receptor, such as and preferably 5-HT2b receptor antagonists;

[0223] - Anti-fibrotic agents, such as 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 as bronchodilators, such as and preferably 5-HT2b receptor antagonists; β2-adrenergic agonists (short- and long-acting), anticholinergics, and theophylline;

[0225] - Agents as antagonists of cytokines and chemokines, such as 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 integrin;

[0226] - Organic nitrates and NO donors, such as and preferably sodium nitroprusside, nitroglycerin, isosorbide mononitrate, isosorbide dinitrate, molsidomine, or SIN-1, as well as inhaled NO;

[0227] - NO-independent but heme-dependent stimulators of soluble guanylate cyclase, such as and preferably the compounds described in WO 00 / 06568, WO 00 / 06569, WO 02 / 42301, WO 03 / 095451, WO 2011 / 147809, WO 2012 / 004258, WO2012 / 028647, and WO 2012 / 059549;

[0228] - NO - independent and heme - independent activators of soluble guanylate cyclase, such as 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] - Reagents that stimulate cGMP synthesis, such as sGC modulators, such as and preferably riociguat, cinaciguat, vericiguat;

[0230] - Prostacyclin analogs or IP receptor agonists, such as and preferably iloprost, beraprost, treprostinil, epoprostenol or Selexipag;

[0231] - Endothelin receptor antagonists, such as and preferably bosentan, darusentan, Ambrisentan or sitaxentan;

[0232] - Reagents that inhibit soluble epoxide hydrolase (sEH), such as and preferably N,N′ - dicyclohexylurea, 12 - (3 - adamantan - 1 - yl - ureido) - dodecanoic acid or 1 - adamantan - 1 - yl - 3 - {5 - [2 - (2 - ethoxyethoxy)ethoxy]pentyl} - urea;

[0233] - Reagents that interact with glucose metabolism, such as and preferably insulin, biguanides, thiazolidinediones, sulfonylureas, acarbose, DPP4 inhibitors, GLP - 1 analogs or SGLT - 1 inhibitors;

[0234] - Natriuretic peptides, such as and preferably atrial natriuretic peptide (ANP, carperitide), brain natriuretic peptide (BNP, nesiritide), C - type natriuretic peptide (CNP) or urodilatin;

[0235] - Natriuretic peptide derivatives, such as and preferably vosoritide, cenderitide, PL3994;

[0236] - Cardiac myosin activators, such as and preferably omecamtiv mecarbil (CK - 1827452);

[0237] - Calcium sensitizers, such as and preferably levosimendan;

[0238] - Agents that affect cardiac energy metabolism, such as and preferably etomoxir, dichloroacetate, ranolazine or trimetazidine, all or part of adenosine A1 receptor agonists, such as GS-9667 (formerly known as CVT-3619), capadenoson and neladenoson;

[0239] - Agents that affect heart rate, such as and preferably ivabradine.

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

[0241] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a platelet aggregation inhibitor, such as and preferably aspirin, clopidogrel, prasugrel, ticagrelor, ticlopidine or dipyridamole.

[0242] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a thrombin inhibitor, such as and preferably ximelagatran, dabigatran, melagatran, bivalirudin or clexane.

[0243] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a GPIIb / IIIa antagonist, such as and preferably tirofiban or abciximab.

[0244] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a factor Xa inhibitor, such as 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, the antibody or fragment thereof of the present invention is administered in combination with heparin or low molecular weight (LMW) heparin derivatives.

[0246] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a vitamin K antagonist, such as and preferably coumarin.

[0247] The antihypertensive agent is preferably understood to be a compound from calcium antagonists, angiotensin AlI antagonists, ACE inhibitors, endothelin antagonists, renin inhibitors, α-blockers, β-blockers, mineralocorticoid receptor antagonists, rho kinase inhibitors, and diuretics.

[0248] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a calcium antagonist, such as and preferably nifedipine, amlodipine, verapamil, or diltiazem.

[0249] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an α-1 blocker, such as and preferably prazosin.

[0250] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a β-blocker, such as and preferably propranolol, atenolol, timolol, indolol, alprenolol, oxprenolol, penbutolol, bunitrolol, metipranolol, nadolol, mepindolol, carteolol, sotalol, betaxolol, celiprolol, bisoprolol, carteolol, esmolol, labetalol, carvedilol, adalimumab, landiolol, nebivolol, epanolol, or bucindolol.

[0251] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an angiotensin AII antagonist, such as and preferably losartan, candesartan, valsartan, telmisartan, or eprosartan, or a dual angiotensin AII antagonist / neprilysin inhibitor, such as and preferably LCZ696 (valsartan / sacubitril).

[0252] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an ACE inhibitor, such as and preferably enalapril, captopril, lisinopril, ramipril, delapril, fosinopril, quinapril, perindopril, or trandolapril.

[0253] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an endothelin antagonist, such as and preferably bosentan, darusentan, ambrisentan, or sitaxentan.

[0254] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a renin inhibitor, such as and preferably aliskiren, SPP-600, or SPP-800.

[0255] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a mineralocorticoid receptor antagonist, such as and preferably finerenone, spironolactone or eplerenone.

[0256] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a loop diuretic, such as furosemide, torasemide, bumetanide and piretanide, with a potassium-sparing diuretic such as amiloride and triamterene, with an aldosterone antagonist, such as spironolactone, canrenoate potassium and eplerenone, and with a thiazide diuretic, such as hydrochlorothiazide, chlorthalidone, xipamide and indapamide.

[0257] Lipid metabolism modifiers are preferably understood as compounds from CETP inhibitors, thyroid receptor agonists, cholesterol synthesis inhibitors such as HMG-CoA reductase inhibitors or squalene synthesis inhibitors, ACAT inhibitors, MTP inhibitors, PPAR-α, PPAR-γ and / or PPAR-δ agonists, cholesterol absorption inhibitors, polymeric bile acid adsorbents, bile acid reabsorption inhibitors, lipase inhibitors and lipoprotein(a) antagonists.

[0258] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a CETP inhibitor, such as and preferably dalcetrapib, anacetrapib, torcetrapib (CP-529414), JJT-705 or CETP vaccine (Avant).

[0259] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a thyroid receptor agonist, such as and preferably D-thyroxine, 3,5,3′-triiodothyronine (T3), CGS23425 or axitirole (CGS 26214).

[0260] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an HMG-CoA reductase inhibitor from statins, such as and preferably lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin, rosuvastatin or pitavastatin.

[0261] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a squalene synthesis inhibitor, such as and preferably BMS-188494 or TAK-475.

[0262] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an ACAT inhibitor, such as and preferably avasimibe, linoleic acid amide, pactimibe, eflucimibe or SMP-797.

[0263] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an MTP inhibitor, such as and preferably impetapide, BMS-201038, R-103757 or JTT-130.

[0264] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a PPAR-γ agonist, such as and preferably pioglitazone or rosiglitazone.

[0265] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a PPAR-δ agonist, such as and preferably GW 501516 or BAY 68-5042.

[0266] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a cholesterol absorption inhibitor, such as and preferably ezetimibe, tiquian or pamaquine.

[0267] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a lipase inhibitor, such as and preferably orlistat.

[0268] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a polymeric bile acid sequestrant, such as and preferably cholestyramine, colestipol, colesolvam, colloid or colestimide.

[0269] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a bile acid reabsorption inhibitor, such as 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 antibody or fragment thereof of the present invention is administered in combination with a lipoprotein(a) antagonist, such as and preferably gemcabene calcium (CI-1027) or niacin.

[0271] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a lipoprotein(a) antagonist, such as and preferably gemcabene calcium (CI-1027) or niacin.

[0272] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an sGC modulator, such as and preferably riociguat, cinaciguat or vericiguat.

[0273] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an agent that affects glucose metabolism, such as and preferably insulin, sulfonylurea, acarbose, DPP4 inhibitor, GLP-1 analog, or SGLT-1 inhibitor.

[0274] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a TGFβ antagonist, such as and preferably pirfenidone, nintedanib, or fresolimumab.

[0275] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a CCR2 antagonist, such as and preferably CCX-140.

[0276] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a TGFα antagonist, such as and preferably adalimumab.

[0277] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a galectin-3 inhibitor, such as and preferably GCS-100.

[0278] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an Nrf-2 inhibitor, such as and preferably bardoxolone.

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

[0280] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a NOX1 / 4 inhibitor, such as and preferably GKT-137831.

[0281] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an agent that affects vitamin D metabolism, such as and preferably calcitriol, alfacalcidol, doxercalciferol, maxacalcitol, paricalcitol, cholecalciferol, or paricalcitol.

[0282] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a cell growth inhibitor, such as and preferably cyclophosphamide.

[0283] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an immunosuppressant, such as and preferably cyclosporine.

[0284] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a phosphate binder, such as and preferably colestilan, sevelamer hydrochloride and sevelamer carbonate, lanthanum and lanthanum carbonate.

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

[0286] In a specific embodiment, the antibody or fragment thereof of the present invention is co-administered with a calcimimetic for the treatment of hyperparathyroidism.

[0287] In a specific embodiment, the antibody or fragment thereof of the present invention is co-administered with a reagent for the treatment of iron deficiency, such as and preferably an iron product.

[0288] In a specific embodiment, the antibody or fragment thereof of the present invention is co-administered with a reagent for the treatment of hyperurikaemia, such as and preferably allopurinol or rasburicase.

[0289] In a specific embodiment, the antibody or fragment thereof of the present invention is co-administered with a glycoprotein hormone for the treatment of anaemia, such as and preferably erythropoietin.

[0290] In a specific embodiment, the antibody or fragment thereof of the present invention is co-administered with a biological product for immunotherapy, such as and preferably abatacept, rituximab, eculizumab or belimumab.

[0291] In a specific embodiment, the antibody or fragment thereof of the present invention is co-administered with a vasopressin antagonist (vaptanes) for the treatment of heart failure, such as and preferably tolvaptan, conivaptan, lixivaptan, mozavaptan, satavaptan or relcovaptan.

[0292] In a specific embodiment, the antibody or fragment thereof of the present invention is co-administered with a Jak inhibitor, such as and preferably ruxolitinib, tofacitinib, baricitinib, CYT387, GSK2586184, lestaurtinib, pacritinib (SB1518) or TG101348.

[0293] In a specific embodiment, the antibody or fragment thereof of the present invention is co-administered with a prostacyclin analogue for the treatment of microthrombi.

[0294] In a specific embodiment, the antibody or fragment thereof of the present invention is co-administered with an alkali treatment, such as and preferably sodium bicarbonate.

[0295] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an mTOR inhibitor, such as and preferably everolimus or rapamycin.

[0296] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an NHE3 inhibitor, such as and preferably AZD1722 or tenapanor.

[0297] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with an eNOS modulator, such as and preferably sapropterin.

[0298] In a specific embodiment, the antibody or fragment thereof of the present invention is administered in combination with a CTGF inhibitor, such as and preferably FG-3019.

[0299] The antibody or fragment thereof for use in the treatment methods of the present invention can be formulated in any conventional manner using one or more physiologically acceptable carriers or excipients. The antibody or fragment thereof of the present invention can be administered by any suitable means, which can vary depending on the type of disorder to be treated. Possible routes of administration include enteral (e.g., oral), parenteral (such as intravenous, intra-arterial, intraperitoneal, intramuscular, subcutaneous, intracardiac, intraventricular, intrathecal, intramedullary, intralesional), pulmonary, and intranasal administration. In addition, the antibody or fragment thereof of the present invention can be administered by infusion, for example, at a reduced dose of the antibody or fragment thereof. Administration is preferably by injection, most preferably by intravenous injection or subcutaneous injection, in part depending on whether the condition is acute or chronic. The amount to be administered will depend on a number of factors, such as the individual's clinical symptoms, sex, age, and / or weight, whether other medications are being given, etc. Those skilled in the art will recognize that the route of administration will vary depending on the disorder or condition to be treated.

[0300] Parenteral delivery methods include topical, intra-arterial, intratumoral, intramuscular, subcutaneous, intramedullary, intrathecal, intraventricular, intravenous, intraperitoneal, or intranasal administration.

[0301] In a specific embodiment, the treatment method includes single or multiple administrations of the antibody or fragment thereof or a pharmaceutical composition comprising the antibody or fragment thereof. The single-dose administrations can be the same or different. Specifically, the treatment method includes 1, 2, 3, 4, 5, or 6 administrations of the antibody or fragment thereof of the present invention, preferably where multiple administrations occur within one to six consecutive months. The antibody or fragment thereof of the present invention can be administered, for example, every 3 to 4 days, weekly, every two weeks, or every three weeks, depending on its half-life and clearance rate. Sequence Listing <110> Bayer AG <120> Atrial Natriuretic Peptide Grafted Antibody <130> CP1200755P <160> 447 <170> PatentIn version 3.5 <210> 1 <211> 10 <212> PRT <213> Artificial <220> <223> Linker 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> Linker 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> Linker sequence <400> 3 Ser Asn Thr Lys Val Asp Lys Lys Val 1 5 <210> 4 <211> 10 <212> PRT <213> Artificial <220> <223> Linker 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> Linker sequence <400> 5 Asp Arg Phe Ser Gly Ser Lys Ser Gly 1 5 <210> 6 <211> 15 <212> PRT <213> Artificial <220> <223> Linker 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> Linker 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> Linker 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> Linker 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> Linker 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> Linker 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> Linker 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> Linker sequence <400> 13 Ala Val Val Asn Val Arg Ala 1 5 <210> 14 <211> 7 <212> PRT <213> Artificial <220> <223> Linker sequence <400> 14 Lys Gly Asp Lys Ile Ala Ile 1 5 <210> 15 <211> 10 <212> PRT <213> Artificial <220> <223> Linker 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> Linker 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> Linker sequence <400> 17 Gln Gly Ile Ile Asn Phe Glu Gln Lys 1 5 <210> 18 <211> 11 <212> PRT <213> Artificial <220> <223> Linker 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> Linker sequence <400> 19 Tyr Glu Lys Glu Lys Glu Lys 1 5 <210> 20 <211> 10 <212> PRT <213> Artificial <220> <223> Linker sequence <400> 20 Gly Gly Ser Thr Pro Leu Lys Ser Leu Ala 1 5 10 <210> 21 <211> 14 <212> PRT <213> Artificial <220> <223> Linker sequence <400> 21 Gly Ile Thr Gly Thr Lys Lys Tyr Gln Ser Ser Pro Asp Gly 1 5 10 <210> 22 <211> 17 <212> PRT <213> Artificial <220> <223> Linker sequence <400> 22 Ser Tyr Ser Tyr Thr Tyr Asn Tyr Ala Glu Tyr Phe Gln His Trp Gly 1 5 10 15 Gln <210> 23 <211> 28 <212> PRT <213> Human <400> 23 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> 24 <211> 28 <212> PRT <213> Human <400> 24 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> 22 <212> PRT <213> Human <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> 23 <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 (without 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 ref 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 <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 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 70 75 80 Asp Arg Ile Gly Ala Gln Ser Gly Leu Gly Cys Asn Ser Phe Arg Tyr 85 90 95 Gly Gly Ser Thr Pro Leu Lys Ser Leu Ala Ser Thr Tyr Tyr Ala Asp 100 105 110 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 115 120 125 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 130 135 140 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> NP-grafted LC <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 variant <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> 21 <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> 24 <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 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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 22 <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> 23 <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> 21 <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> 23 <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> 21 <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> 23 <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> 21 <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> 23 <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> 23 <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> 23 <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> 23 <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> 21 <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> 22 <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> 22 <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> 22 <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> 24 <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 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> 21 <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 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> 21 <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> 21 <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> 21 <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> 21 <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> 22 <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> 22 <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> 22 <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> 22 <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> 22 <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> 23 <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> 24 <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> 21 <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> 23 <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> 22 <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> 24 <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> 21 <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> 21 <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> 24 <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> 23 <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> 22 <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 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 Val Val Val Thr Ser His Gln Ala Pro Gly 50 55 60 Gly Gly Ser Gly Gly Met Val Gln Gly Ser Gly Cys Phe Gly Arg Lys 65 70 75 80 Met Asp Arg Ile Ser Ser Ser Ser Gly Leu Gly Cys Lys Val Leu Arg 85 90 95 Arg Gly Glu Lys Lys Lys Leu Lys Ser Leu Ala Tyr 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> 444 <211> 499 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 444 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 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 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> 445 <211> 499 <212> PRT <213> Artificial <220> <223> NP-grafted HC <400> 445 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 Tyr Lys 100 105 110 Gly Ala Asn...

Claims

1. An antibody or fragment thereof, comprising a heterologous amino acid sequence, wherein the heterologous amino acid sequence consists of an N-terminal linker sequence (Ntls), atrial natriuretic peptide (ANP), and a C-terminal linker sequence (Ctls), wherein (i) the light chain of the antibody or fragment thereof comprises the amino acid sequence SEQ ID NO 66, and the heavy chain of the antibody or fragment thereof comprises an amino acid sequence selected from any one of SEQ ID NOs 67 to 79; or (ii) the heavy chain of the antibody or fragment thereof comprises the amino acid sequence SEQ ID NO 65, the light chain of the antibody or fragment thereof comprises an amino acid sequence selected from any one of SEQ ID NOs 80 and 81, and wherein the antibody fragment is selected from Fab, Fab’, Fab’-SH, F(ab’)2, and Fv fragments; bivalent antibodies; single domain antibodies (Dab); linear antibodies; single-chain antibody molecules (ScFv); and disulfide-stabilized Fv antibody fragments (dsFv).

2. An antibody or fragment thereof, comprising a heterologous amino acid sequence, wherein the heterologous amino acid sequence consists of an N-terminal linker sequence (Ntls), atrial natriuretic peptide (ANP), and a C-terminal linker sequence (Ctls), wherein (i) the light chain of the antibody or fragment thereof consists of the amino acid sequence SEQ ID NO 66, and the heavy chain of the antibody or fragment thereof consists of an amino acid sequence selected from any one of SEQ ID NOs 67 to 79; or (ii) the heavy chain of the antibody or fragment thereof consists of the amino acid sequence SEQ ID NO 65, the light chain of the antibody or fragment thereof consists of an amino acid sequence selected from any one of SEQ ID NOs 80 and 81, and wherein the antibody fragment is selected from Fab, Fab’, Fab’-SH, F(ab’)2, and Fv fragments; bivalent antibodies; single domain antibodies (Dab); linear antibodies; single-chain antibody molecules (ScFv); and disulfide-stabilized Fv antibody fragments (dsFv).

3. The antibody or fragment thereof according to claim 1, wherein the light chain comprises the amino acid sequence SEQ ID NO 66, and the heavy chain comprises an amino acid sequence selected from any one of SEQ ID NOs 67 to 79.

4. The antibody or fragment thereof according to claim 2, wherein the light chain consists of the amino acid sequence SEQ ID NO 66, and the heavy chain consists of an amino acid sequence selected from any one of SEQ ID NOs 67 to 79.

5. The antibody or fragment thereof according to claim 3, wherein the light chain comprises the amino acid sequence SEQ ID NO 66, and the heavy chain comprises the amino acid sequence SEQ ID NO 76.

6. The antibody or fragment thereof according to claim 4, wherein the light chain consists of the amino acid sequence SEQ ID NO 66, and the heavy chain consists of the amino acid sequence SEQ ID NO 76.

7. A composition, comprising the antibody or fragment thereof according to any one of claims 1 to 6 and optionally a pharmaceutically acceptable carrier compound.

8. A nucleic acid or nucleic acid mixture encoding an antibody or fragment thereof according to any one of claims 1 to 6.

9. A host cell comprising the nucleic acid or nucleic acid mixture according to claim 8.

10. A method of producing an antibody or fragment thereof, comprising culturing the host cell according to claim 9 under conditions suitable for expressing the antibody or fragment thereof.

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