Anti-DPP3 antibody directed to and binding to DPP3 for the treatment of shock in extreme-critically ill patients

Abstract

The present invention relates to an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said patient is an extreme-critically ill patient. In certain aspects, the present invention relates to an anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, wherein said shock patient has acute kidney injury (AKI).

Description

[0001] T75380WO BOEHMERT & BOEHMERT

[0002] Anti-DPP3 antibody directed to and binding to DPP3 for the treatment of shock in extreme- critically ill patients

[0003] FIELD OF THE INVENTION

[0004] Subject matter of the invention is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said patient is an extreme-critically ill patient.

[0005] BACKGROUND

[0006] Shock is characterized by decreased oxygen delivery and / or increased oxygen consumption or inadequate oxygen utilization leading to cellular and tissue hypoxia. It is a life-threatening condition of circulatory failure and most commonly manifested as hypotension (systolic blood pressure less than 90 mm Hg or a mean arterial pressure (MAP) less than 65 mmHg). Shock is divided into four main types based on the underlying cause: hypovolemic, obstructive, cardiogenic, and distributive shock (Vincent and De Backer 2014. N. Engl. J. Med. 370(6): 583).

[0007] Hypovolemic shock is characterized by decreased intravascular volume and can be divided into two broad subtypes: hemorrhagic and non-hemorrhagic. Common causes of hemorrhagic hypovolemic shock include gastrointestinal bleed, trauma, vascular etiologies (e.g. ruptured abdominal aortic aneurysm, tumor eroding into a major blood vessel), and spontaneous bleeding in the setting of anticoagulant use. Common causes of non-hemorrhagic hypovolemic shock include vomiting, diarrhea, renal loss, skin losses / insensible losses (e.g. bums, heat stroke), or third-space loss in the setting of pancreatitis, cirrhosis, intestinal obstruction, trauma. For review see Koya and Paul 2018. Shock. StatP earls [Internet], Treasure Island (FL): StatP earls Publishing: 2019-2018 Oct 27.

[0008] Obstructive shock is due to a physical obstruction of the great vessels (i.e., the large arteries or veins that connect directly to the heart and carry blood to or from it) or the heart itself. Several conditions can result in this form of shock (e.g. cardiac tamponade, tension pneumothorax, pulmonary embolism, aortic stenosis). For review see Koya and Paul 2018. Shock. StatPearls [Internet], Treasure Island (FL): StatP earls Publishing: 2019-2018 Oct 27.

[0009] According to the cause, there are four types of distributive shock: neurogenic shock (decreased sympathetic stimulation leading to decreased vasal tone), anaphylactic shock, septic shock, and shock due to adrenal crisis. In addition to sepsis, distributive shock can be caused by systemic inflammatory response syndrome (SIRS) due to conditions other than infection such as pancreatitis, bums, or trauma. Other causes include toxic shock syndrome (TSS), anaphylaxis (a sudden, severe allergic reaction), adrenal insufficiency (acute worsening of chronic adrenal insufficiency, destmction or removal of the adrenal glands, suppression of adrenal gland function due to exogenous steroids, hypopituitarism and metabolic failure of hormone production), reactions to drugs or toxins, heavy metal poisoning, hepatic (liver) insufficiency, and damage to the central nervous system. For review see Koya and Paul 2018.

[0010] Shock. StatP earls [Internet], Treasure Island (FL): StatPearls Publishing: 2019-2018 Oct 27.

[0011] Cardiogenic shock (CS) is a complete breakdown of the circulatory system post an acute event such as acute myocardial infarction (AMI), acute decompensated heart failure (HF), or septic cardiomyopathy and results in systemic hypoperfusion, coronary ischemia, increased ventricular fdling pressures, inflammation, multiorgan failure, and often death. AMI with subsequent ventricular dysfunction is the most frequent cause of CS accounting for approximately 80% of cases. Mechanical complications such as ventricular septal (4%) or free wall rupture (2%), and acute severe mitral regurgitation (7%) are less frequent causes of CS after AMI. (Hochman et al. 2000. J Am Coll Cardiol 36: 1063-1070}. Non-AMI- related CS may be caused by decompensated valvular heart disease, acute myocarditis, arrhythmias, etc. with heterogeneous treatment options.

[0012] Septic shock is a potentially fatal medical condition that occurs when sepsis, which is organ injury or damage in response to infection, leads to dangerously low blood pressure and abnormalities in cellular metabolism. The Third International Consensus Definitions for Sepsis and Septic Shock (Sepsis-3) defines septic shock as a subset of sepsis in which particularly profound circulatory, cellular, and metabolic abnormalities are associated with a greater risk of mortality than with sepsis alone (Singer et al. 2016. JAMA. 315 (8): 801-10). Patients with septic shock can be clinically identified by a vasopressor requirement to maintain a mean arterial pressure of 65 mm Hg or greater and serum lactate level greater than 2 mmol / L (>18 mg / dL) in the absence of hypovolemia. This combination is associated with hospital mortality rates greater than 40% (Singer et al. 2016. JAMA. 315 (8): 801-10). The primary infection is most commonly caused by bacteria, but also may be by fungi, viruses, or parasites. It may be located in any part of the body, but most commonly in the lungs, brain, urinary tract, skin, or abdominal organs. It can cause multiple organ dysfunction syndrome (formerly known as multiple organ failure) and death. Frequently, people with septic shock are cared for in intensive care units. It most commonly affects children, immunocompromised individuals, and the elderly, as their immune systems cannot deal with infection as effectively as those of healthy adults. The mortality rate from septic shock is approximately 25-50%.

[0013] CS is defined as a state of critical end organ hypoperfusion due to reduced cardiac output. Notably, CS forms a spectrum that ranges from mild hypoperfusion to profound shock. Criteria for the diagnosis of CS are: (i) systolic blood pressure, <90 mmHg for >30 min or vasopressors required to achieve a blood pressure >90 mmHg; (ii) pulmonary congestion or elevated left-ventricular filling pressures; (iii) signs of impaired organ perfusion with at least one of the following criteria: (a) altered mental status; (b) cold, clammy skin; (c) oliguria (< 0.5 mL / kg / h or <30 mL / h); (d) increased serum-lactate (Reynolds and Hochman 2008. Circulation 117: 686-697}. Around 150,000 patients in the US, and 250,000 in Europe suffer from CS per year. The prognosis of CS, despite best standard of care (SOC), remains very poor, with up to 50% 30-day mortality (Muzafarova et al. 2023), and no effective, causal treatment options available (Arrigo et al. 2023)(Arrigo et al. 2021).

[0014] The current SOC for CS is largely supportive, focused on providing hemodynamic support (Thiele et al. 2012)(Van Diepen et al. 2017)(Aissaoui et al. 2020)(Thiele et al. 2023). Current CS management encompasses a mix of pharmacological and mechanical interventions including implantation of mechanical circulatory support devices (MCS) and organ replacement therapies for secondary organ failure (such as dialysis and mechanical ventilation) (Chioncel et al. 2020). However, even with optimal treatment following best practice guidelines, many CS patients still do not survive or suffer significant complications and functional limitations (Van Diepen et al. 2017), as none of the currently available interventions, neither pharmaceutical nor mechanical, are addressing the underlying pathophysiological pathways leading to CS.

[0015] All forms of shock can become refractory, as evidenced by unresponsiveness to high-dose vasopressors (Udupa and Shetty 2018. Indian J Respir Care 7: 67-72}. In general, refractory shock has been defined as requirement of noradrenaline infusion of >0.5 pg / kg / min despite adequate volume resuscitation. Mortality in these patients may be as high as 94% and the assessment and management of these patients requires a much more aggressive approach for survival. The term “refractory shock” is used when the tissue perfusion cannot be restored with the initial corrective measures employed (e.g. vasopressors) and may therefore be referred to as “high vasopressor-dependent” or “vasopressor-resistant” shock (Udupa and Shetty 2018. Indian J Respir Care 7: 67-72}.

[0016] Dipeptidyl peptidase 3 - also known as Dipeptidyl aminopeptidase III, Dipeptidyl arylamidase III, Dipeptidyl peptidase III, Enkephalinase B, or red cell angiotensinase; short name: DPP3, DPPIII - is a metallopeptidase that removes dipeptides from physiologically active peptides, such as enkephalins and angiotensins. DPP3 was identified and its activity measured in extracts of purified bovine anterior pituitary (Ellis and Nuenke 1967). The enzyme, which is listed as EC 3.4. 14.4, has a molecular mass of about 83 kDa and is highly conserved in procaryotes and eucaryotes (Prajapati and Chauhan 2011). The amino acid sequence of the human variant is depicted in SEQ ID NO: 1. DPP3 is a mainly cytosolic peptidase which is ubiquitously expressed. Despite lacking a signal sequence, a few studies reported membranous activity (Lee and Snyder 1982).

[0017] DPP3 is a zinc-depending exo-peptidase belonging to the peptidase family M49. It has a broad substrate specificity for oligopeptides from three / four to ten amino acids of various compositions and is also capable of cleaving after proline. DPP3 is known to hydrolyze dipeptides from the N-terminus of its substrates, including angiotensin II, III, and IV; Leu- and Met-enkephalin; endomorphin 1 and 2. The metallopeptidase DPP3 has its activity optimum at pH 8.0-9.0 and can be activated by addition of divalent metal ions, such as Co2+and Mg2+.

[0018] Structural analysis of DPP3 revealed the catalytic motifs HELLGH (human DPP3 [hDPP3] 450-455; SEQ ID NO: 14) and EECRAE (hDPP3 507-512; SEQ ID NO: 15), as well as the following amino acids, that are important for substrate binding and hydrolysis: Glu316, Tyr, 318, Asp366, Asn391, Asn394, His568, Arg572, Arg577, Lys666, and Arg669 (Prajapati and Chauhan 2011) (Kumar et al. 2016); numbering refers to the sequence of human DPP3, see SEQ ID NO: 1). Considering all known amino acids or sequence regions that are involved in substrate binding and hydrolysis, the active site of human DPP3 can be defined as the area between amino acids 316 and 669.

[0019] The most prominent substrate of DPP3 is angiotensin II (Ang II), the main effector of the reninangiotensin system (RAS). The RAS is activated in cardiovascular diseases (Dostal et al. 1997) (Roks et al. 1997), sepsis, and septic shock (Correa, Takala, and Jakob 2015). Ang II, in particular, has been shown to modulate many cardiovascular functions including the control of blood pressure and cardiac remodeling.

[0020] Recently, two assays were generated, characterized, and validated to specifically detect DPP3 in human bodily fluids (e.g., blood, plasma, serum): a luminescence immunoassay (LIA) to detect DPP3 protein concentration and an enzyme capture activity assay (ECA) to detect specific DPP3 activity (Rehfeld et al., 2019 JALM 3(6): 943-953). A washing step removes all interfering substances before the actual detection of DPP3 activity is performed. Both methods are highly specific and allow the reproducible detection of DPP3 in blood samples.

[0021] Additionally, the kit sphingotest® DPP3 (Sphingotec GmbH, Hennigsdorf, Germany) is a commercially available immunoluminometric assay (ILMA) for the quantitative measurement of dipeptidyl peptidase 3 in human EDTA plasma. It is based on the LIA method described in Rehfeld et al. 2019 whereby it uses the two identified antibodies disclosed in Rehfeld (Rehfeld et al. 2019). It is a sandwich ELISA using two antigen-specific monoclonal antibodies. The first antibody is the capture antibody binding DPP3 to a microtiter plate well and the other is the detected tracer antibody binding DPP3 at a different epitope and is labeled with a luminescent label.

[0022] Circulating DPP3 (cDPP3) levels were shown to be increased in septic, cardiogenic, and vasodilatory shock patients (Rehfeld et al. 2019). Moreover, an increased cDPP3 level was associated with an increased risk of short-term mortality and severe organ dysfunction in patients with cardiogenic shock (Deniau et al. 2020). Moreover, patients with severe sepsis or septic shock showed that the higher the initial cDPP3 was, the greater the need for organ support and vasopressors upon admission and the longer the need for vasopressor(s), mechanical ventilation, or renal replacement therapy (RRT) and the higher the need for fluid load (Blet et al. 2021).

[0023] WO2017 / 182561 describes methods for determining the total amount or active DPP3 in a sample of a patient for the diagnosis of a disease related to necrotic processes. It further describes a method of treatment of necrosis-related diseases by antibodies directed to DPP3.

[0024] WO2019 / 081595 describes DPP3 binders directed to and binding to specific DPP3 epitopes and its use in the prevention or treatment of diseases that are associated with oxidative stress.

[0025] WO2021 / 185786 describes methods for determining DPP3 in a sample of a patient for the diagnosis, risk prediction, prognosis, and monitoring in a patient infected with a coronavirus. It further describes an inhibitor of the activity of DPP3 for use in therapy or intervention in a patient infected.

[0026] WO2024 / 126793 describes an inhibitor of the activity of DPP3 for use in therapy or intervention in a critically ill patient with a reduction in pulmonary function for the improvement of pulmonary function.

[0027] Thus, efforts have been made to pharmacologically inhibit supranormal concentrations of cDPP3.

[0028] Numerous binders that inhibit DPP3 have been described (Marija Abramic and Agic 2022): Except for the polypeptide aprotinin, all others are small molecules and include flavonoids, coumarin, and benzimidazole derivatives. Presented are current strategies for the discovery or development of DPP3 inhibitors, and mechanisms of inhibitory actions. The most potent inhibitors yet reported (propioxatin A and B, Tyr-Phe- and Phe-Phe-NHOH, and JMV-390) are active in low nanomolar range and contain a hydroxamic acid moiety. Oligopeptides from the hemorphin group, valorphin and tynorphin, exhibit high inhibitory potential but which are poor substrates of DPP3.

[0029] An anti-DPP3 antibody, particularly a humanized anti-DPP3 antibody, and more particularlya humanized monoclonal IgGl antibody, specifically binds to cDPP3, thereby targeting and modulating the activity of cDPP3. Its mode of action is relevant in acute diseases that are associated with massive cell death and uncontrolled release of intracellular DPP3 into the bloodstream. Translocated DPP3 remains active in the circulation where it cleaves bioactive peptides in an uncontrolled manner.

[0030] A humanized anti-DPP3 antibody is able to inhibit cDPP3 thereby inhibiting degradation of bioactive peptides in the bloodstream. This inhibition results in stabilization of cardiovascular and renal function and reduction of short-term mortality. Preclinical studies of humanized anti-DPP3 antibody in animal models of cardiovascular failure showed impressive and instant efficacy. In several preclinical cardiovascular failure models, humanized anti-DPP3 antibody has shown to normalize ejection fraction and kidney function and reduces mortality (Malovan et al. 2023).

[0031] It was the surprising finding of the present invention that an anti-DPP3 antibody or fragment thereof binding to DPP3 could be used for the treatment of shock, in an extreme-critically ill patient having shock.

[0032] The object of the present invention is the provision of an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said patient is an extreme-critically ill patient. A specific finding of the present invention is that an anti-DPP3 antibody could be used for the treatment of shock in an extreme-critically ill patient having shock, wherein said patient has cardiogenic shock or combined cardiogenic and septic shock.

[0033] Further object of the present invention is the provision of an anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient having shock for improvement or stabilization of kidney function, wherein said shock patient has acute kidney injury (AKI).

[0034] SUBJECT MATTER OF THE INVENTION

[0035] Subject matter of the present invention is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, wherein said patient is an extreme-critically ill patient.

[0036] Further subject matter of the present invention is an anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient having shock for improvement or stabilization of kidney function, wherein said shock patient has acute kidney injury (AKI).

[0037] Further subject matter of the present invention is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, wherein said patient is an extreme-critically ill patient, wherein said extreme-critically ill patient is characterized by a level of DPP3 in a sample of a bodily fluid of said patient above a threshold, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and wherein the level of DPP3 is determined by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3. Further subject matter of the present invention is a method comprising determining the level of DPP3 in a sample of a bodily fluid of a patient having shock, wherein said patient is an extreme-critically ill patient, by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and, if said level of DPP3 in said sample is above a threshold, said patient is requiring administration of an anti-DPP3 antibody or fragment thereof for the treatment of said shock.

[0038] Further subject matter of the present invention is a method for the treatment of shock in an extreme- critically ill patient having shock, comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof, wherein said extreme-critically ill patient is characterized by a level of DPP3 in a sample of a bodily fluid of said patient above a threshold, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and wherein the level of DPP3 is determined by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3.

[0039] Further subject matter of the present invention is a method for the treatment of shock in an extreme- critically ill patient having shock, comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof, wherein the level of DPP3 determined in a sample of a bodily fluid of said patient by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3 is above a threshold, and wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample.

[0040] Further subject matter of the present invention is a method comprising determining the level of DPP3 in a sample of a bodily fluid of an extreme-critically ill patient having shock by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and, if said level of DPP3 in said sample is above a threshold, administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof.

[0041] In one embodiment of the invention said shock is selected from the group comprising shock due to hypovolemia, cardiogenic shock, obstructive shock, and distributive shock. In one embodiment of the present invention hypovolemic shock is hemorrhagic and non-hemorrhagic shock.

[0042] In a further embodiment, hypotension in a shock patient is defined as having a systolic blood pressure less than 90 mm Hg or MAP less than 65 mmHg. In one embodiment of the present invention, criteria for the diagnosis of CS are: (i) systolic blood pressure, <90 mmHg for >30 min or vasopressors required to achieve a blood pressure >90 mmHg; (ii) pulmonary congestion or elevated left-ventricular filling pressures; (iii) signs of impaired organ perfusion with at least one of the following criteria: (a) altered mental status; (b) cold, clammy skin; (c) oliguria (< 0.5 mL / kg / h or <30 mL / h); (d) increased serum -lactate.

[0043] In one embodiment of the present invention, CS is caused by AMI with subsequent ventricular dysfunction, ventricular septal or free wall rupture, acute severe mitral regurgitation, decompensated valvular heart disease, acute myocarditis, and arrhythmias.

[0044] One embodiment of the present invention is an anti DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said shock is cardiogenic shock.

[0045] One embodiment of the present invention is an anti DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said shock is septic shock.

[0046] Another embodiment of the present invention is an anti DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said shock patient has combined cardiogenic and septic shock. In other words, an embodiment of the present invention is an anti DPP3 antibody or fragment thereof binding to DPP 3 for the treatment of shock in a patient having shock, wherein said shock patient is in particular a cardiogenic shock patient that has additionally septic shock.

[0047] One embodiment of the present invention is an anti DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said shock is refractory shock.

[0048] In a preferred embodiment of the invention is an anti DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said shock patient has combined cardiogenic and septic shock and said cardiogenic and / or septic shock is refractory shock.

[0049] In one embodiment of the present invention, patients with refractory shock have one or more features of inadequate perfusion such as hypotension (mean arterial blood pressure <65 mmHg), tachycardia, cold peripheries, prolonged capillary refill time, and tachypnea consequent to the hypoxia and acidosis. Fever may be seen in septic shock.

[0050] In another embodiment of the present invention, patients with refractory shock have one or more other signs of hypoperfusion such as altered sensorium, hyperlactatemia, and oliguria. These well-known signs of shock are not helpful in identifying whether the problem is at the pump (heart) or circuitry (vessels and tissues). In a further embodiment of the present invention, different types of shock coexist in said patient.

[0051] In a specific embodiment of the invention said refractory shock is vasopressor-resistant, that is defined by unresponsiveness of the patient to high dose vasopressors (e.g. >0.5 pg / kg / min noradrenaline, epinephrine, norepinephrine, angiotensin II).

[0052] The term "patient" as used herein refers to a living human that is receiving medical care or that should receive medical care due to a disease. A patient according to the present invention may be a patient suffering from shock, including a patient suffering from hypovolemic, cardiogenic, obstructive, or distributive shock. In one embodiment of the present invention the patient may be suffering from multiple forms of shock, such as a combination of cardiogenic and distributive shock. In a further embodiment of the invention the patient may be suffering from septic shock. In yet another embodiment of the present invention the patient may be suffering from refractory shock.

[0053] “Organ dysfunction” is a condition where an organ does not perform its expected function. Organ dysfunction in the context of the present invention is in particular a dysfunction of the respective organ to such a degree that normal homeostasis cannot be maintained without external clinical intervention. “Multi -organ dysfunction” denotes an organ dysfunction of at least two organs.

[0054] An extreme-critically ill patient according to the present invention is defined in particular as a patient who has a 24-hour probability of mortality of at least 70%, preferably of at least 80%, more preferably of at least 90%, most preferably of at least 95% and / or has organ dysfunction requiring mechanical or pharmacological support of at least two organs.

[0055] In one embodiment of the invention a patient is considered as an extreme-critically ill patient if said patient is considered as having a 24-hour probability of mortality of at least 70%, preferably of at least 80%, more preferably of at least 90%, most preferably of at least 95% and / or has organ dysfunction requiring mechanical or pharmacological support of at least two organs.

[0056] The probability of mortality is in particular predicted by the treating intensive care physician based on his / her overall clinical assessment. It has been demonstrated from observational studies that intensive care physicians discriminate between survivors and non-survivors more accurately than do objective scoring systems in the first 24 hours of ICU admission (Sinuff et al. 2006).

[0057] In one embodiment of the invention said extreme-critically ill patient is characterized by a level of lactate of 2 mmol / 1 or higher and / or a level of IL-6 of 5.9 pg / ml or higher and / or a level of CRP of 1 mg / dL or higher and / or a level of PCT of 0.5 ng / ml or higher and / or a level of bio-ADM of 70 pg / ml or higher and / or serum creatinine (sCr) of 1.18 mg / dL (for men) and 1.02 mg / dL (for women) and / or a supranormal level of NT-proBNP in a sample of bodily fluid of said patient.

[0058] In one embodiment of the invention said organ dysfunction is defined as cardiovascular dysfunction, respiratory dysfunction (lung), kidney dysfunction, liver dysfunction, central nervous system dysfunction, or dysfunction of the coagulation system. As used herein, organ dysfunction denotes a condition or a state of health where an organ does not perform its expected function. “Organ failure” denotes an organ dysfunction to such a degree that normal homeostasis cannot be maintained without external clinical intervention. Said organ failure may pertain to an organ selected from the group comprising kidney, liver, heart, lung, nervous system. By contrast, organ function represents the expected function of the respective organ within physiologic ranges. The person skilled in the art is aware of the respective function of an organ during medical examination.

[0059] Organ dysfunction may be defined by the sequential organ failure assessment score (SOFA-Score) or the components thereof. The SOFA score, previously known as the sepsis-related organ failure assessment score (Singer et al. 2016) is used to track a person's status during the stay in an intensive care unit (ICU) to determine the extent of a person's organ function or rate of failure. The score is based on six different scores, one each for the respiratory, cardiovascular, hepatic, coagulation, renal, and neurological systems each scored from 0 to 4 with an increasing score reflecting worsening organ dysfunction. The criteria for assessment of the SOFA score are described for example in Lamden et al. (Lambden et al. 2019). SOFA score may traditionally be calculated on admission to ICU and at each 24- h period that follows.

[0060] In one embodiment of the invention said organ support is selected from kidney support, respiratory (lung) support, cardiovascular support, and / or liver support.

[0061] In another embodiment respiratory (lung) support is defined as non-invasive, invasive mechanical ventilation, or ECMO.

[0062] Mechanical ventilation or assisted ventilation is the medical term for artificial ventilation where mechanical means are used to assist or replace spontaneous breathing, in one embodiment mechanical ventilation involves a machine called a ventilator or the breathing may be assisted manually by a suitably qualified professional, such as an anesthesiologist, respiratory therapist, Registered Nurse, or paramedic, by compressing a bag valve mask device. Mechanical ventilation is termed "invasive" if it involves any instrument inside the trachea through the mouth. In one embodiment mechanical ventilation is invasive mechanical ventilation comprising an endotracheal tube or a puncture through the skin to gain access to the lungs, such as a tracheostomy tube. In a further embodiment mechanical ventilation is non-invasive ventilation comprising face or nasal masks. In a further embodiment non-invasive ventilation is used in appropriately selected conscious patients.

[0063] Extracorporeal membrane oxygenation (ECMO), also known as extracorporeal life support (ECLS), is an extracorporeal technique for providing prolonged cardiac and respiratory support to persons whose heart and lungs are unable to provide an adequate amount of gas exchange or perfusion to sustain life. The technology for ECMO is largely derived from cardiopulmonary bypass, which provides shorter- term support with arrested native circulation. ECMO works by removing blood from the person's body and artificially removing carbon dioxide from, and adding oxygen to, the patient's red blood cells.

[0064] Pulmonary function is assessed, particularly in patients under invasive ventilation in critical care, with the Horowitz index (synonyms: oxygenation after Horowitz, Horowitz coefficient, P / F ratio). It is useful for evaluating the extent of damage to the lungs or to evaluate lung function. The Horowitz index (PaO2 / FiO2 ratio) is a well-known biomarker of acute pulmonary injury and predicts mortality in patients with acute respiratory failure. In one embodiment, the Horowitz index is defined as the ratio of partial pressure of oxygen in blood (PaO2), in millimeters of mercury, and the fraction of oxygen in the inhaled air (FiO2) - the PaO2 / FiO2 ratio. In healthy lungs the Horowitz index depends on age and usually falls between 350 and 500. In an embodiment of the present invention, the Horowitz index has a value below 300, wherein a Horowitz index of 300 is the threshold for mild lung injury, wherein a Horowitz index of 200 is of the threshold for a moderately severe lung injury and wherein a Horowitz index of below 100 is the threshold for a severe injury.

[0065] In one embodiment said requirement of organ support is respiratory (lung) support, wherein requirement for respiratory (lung) support by non-invasive and invasive mechanical ventilation is defined by a Horovitz Index (P / F ratio) of 200 or below, preferably 150 or below, preferred 100 or below.

[0066] Kidney support is defined as renal replacement therapy (RRT).

[0067] RRT replaces the normal blood-filtering function of the kidneys. Renal replacement therapy may refer to dialysis (e.g. hemodialysis or peritoneal dialysis), hemofiltration, and hemodiafiltration. Such techniques are various ways of diverting the blood into a machine, cleaning it, and then returning it to the body. The hemodialysis, hemofiltration, and hemodiafiltration may be continuous or intermittent and can use an arteriovenous route (in which blood leaves from an artery and returns via a vein) or a venovenous route (in which blood leaves from a vein and returns via a vein). This results in various types of RRT. For example, the RRT may be selected from the group of, but not limited to continuous renal replacement therapy (CRRT), continuous hemodialysis (CHD), continuous arteriovenous hemodialysis (CAVHD), continuous venovenous hemodialysis (CVVHD), continuous hemofiltration (CHF), continuous arteriovenous hemofiltration (CAVH or CAVHF), continuous venovenous hemofiltration (CVVH or CVVHF), continuous hemodiafiltration (CHDF), continuous arteriovenous hemodiafiltration (CAVHDF), continuous venovenous hemodiafiltration (CVVHDF), intermittent renal replacement therapy (IRRT), intermittent hemodialysis (IHD), intermittent venovenous hemodialysis (IWHD), intermittent hemofiltration (IHF), intermittent venovenous hemofiltration (IVVH or IVVHF), intermittent hemodiafiltration (IHDF), and intermittent venovenous hemodiafiltration (IVVHDF).

[0068] In a specific embodiment of the invention renal replacement therapy is selected from the group comprising dialysis (hemodialysis or peritoneal dialysis), hemofiltration, and hemodiafiltration.

[0069] From the definition of the cardiovascular component of the SOFA Score it can be delineated that pharmacological support of the cardiovascular system is required, if the mean arterial pressure decreases below 70 mm Hg. Support is given by administration of vasoactive drugs such as dopamine, epinephrine, or norepinephrine. In severe cardiovascular dysfunction, doses given are dopamine > 15 pg / kg / min OR epinephrine > 0.1 pg / kg / min OR norepinephrine > 0.1 pg / kg / min.

[0070] In one embodiment said requirement of organ support is support of the cardiovascular system, wherein said support of the cardiovascular system is selected from vasoactive drugs such as dopamine, epinephrine, norepinephrine, midrodine, Hydroxocobalamin, Metaraiminol, Methylene blue, Terlipressin, Angiotensin II, phenylephrine, and vasopressin (Kotani et al. 2023).

[0071] Declining liver function can be described by increasing blood concentrations of bilirubin. In severe liver dysfunction, bilirubin can exceed 12 mg / dL. Extracorporeal Liver Support (ECLS) is a system that performs the essential functions of the liver via filtration and detoxification of blood within an external device with the end goal of reducing mortality or to bridge a patient to liver transplant. In some cases, ECLS can temporarily mitigate sequelae of liver failure such as hepatic encephalopathy and multi -organ failure long enough for the native liverto recover function on its own. The two main modalities of ECLS are divided into artificial (MARS, Prometheus, SPAD and HepaWash) and bio-artificial categories (HepatAssist and ELAD) (Nguyen et al. 2022).

[0072] Subject matter of the invention is also an anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, wherein said shock patient has acute kidney injury (AKI). In certain embodiments of the invention relating to an anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient having AKI, said critically ill patient with shock is under renal replacement therapy.

[0073] In certain embodiments of the invention relating to an anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient having AKI, said shock is selected from the group comprising shock due to hypovolemia, cardiogenic shock, obstructive shock and distributive shock.

[0074] In certain embodiments of the invention relating to an anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient having AKI, said patient has combined cardiogenic and septic shock.

[0075] In certain embodiments of the invention relating to an anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient having AKI, said shock is refractory shock. certain embodiments of the invention relating to an anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient having AKI, an anti-DPP3 antibody is used and wherein the heavy chain comprises at least one CDR, preferably at least three CDRs selected from the group comprising:

[0076] SEQ ID No. 7: GFSLSTSGMS,

[0077] SEQ ID No. 8: IWWNDNK and / or

[0078] SEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR, preferably at least three CDRs selected from the group comprising:

[0079] SEQ ID No. 10: RSLVHSIGSTY,

[0080] CDR2: KVS and / or

[0081] SEQ ID No. 11: SQSTHVPWT certain embodiments of the invention relating to an anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient having AKI, said anti- DPP3 antibody comprises the following sequences: or a sequence that is > 80% identical to it, > 90% identical to it, > 95% identical to it, preferably > 98%, preferably > 99%.

[0082] In certain embodiments of the invention relating to an anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient having AKI, said anti-DPP3 antibody comprises the following sequences: SEQ ID No. 34

[0083] QITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEWLAHIWWNDNKSYNP ALKSRLTITRDTSKNQVVLTMTNMDPVDTGTYYCARNYSYDYWGQGTLVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV PSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ

[0084] KSLSLSPG and / or

[0085] SEQ ID No. 35

[0086] DIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLYWYLQKPGQSPQLLIYKVSNRFSGVPD RFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLS SP VTKSFNRGEC or a sequence that is > 80% identical to it, > 90% identical to it, > 95% identical to it, preferably > 98%, preferably > 99%.

[0087] In certain embodiments of the invention relating to an anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient having AKI, said anti-DPP3 antibody is administered at a dose from 1 to 30 mg / kg bodyweight, preferably 2 to 20 mg / kg bodyweight, preferably 3 to 15 mg / kg bodyweight, preferably 5 to 12 mg / kg body weight, preferably 7 to 10 mg / kg bodyweight, more preferably 10 mg / kg bodyweight.

[0088] In certain embodiments of the invention relating to an anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient having AKI, said anti-DPP3 is administered over the course of 1 to 3 hours, preferably 1.5 to 2.5 hours, more preferably 2 hours and / or wherein said anti- DPP3 antibody is administered at a dose of 10 mg / kg bodyweight as infusion over the course of 1 to 3 hours, preferably 1.5 to 2.5 hours, more preferably 2 hours.

[0089] Acute kidney injury (AKI) affects approximately 10-15% of patients admitted to the hospital (Ronco et al. 2019. Acute kidney injury. Lancet 394(10212): 1949-64). In critically ill patients, its incidence has been reported to exceed 50% (Ronco et al. 2019. Acute kidney injury. Lancet 394(10212): 1949-64: Hoste et al. 2015. Epidemiology of acute kidney injury in critically ill patients: the multinational AKI- EPI study. Intensive Care Medicine. 41(8): 1411-23). For example, sepsis is associated with a greater risk of developing AKI (Bellomo et al. Acute kidney injury in sepsis. Intensive Care Med. 2017;43(6):816-28 and the costs associated with AKI management impose a significant burden on healthcare systems worldwide (OstermannM, Cerda J. The Burden of Acute Kidney Injury and Related Financial Issues. Contrih Nephrol. 2018;193:100-12). The negative consequences of AKI extend beyond the acute phase, with progression to acute kidney disease (no recovery from AKI after seven days) and chronic kidney disease (CKD) (entering CKD after recovery from AKI) (Chawla et al. The severity of acute kidney injury predicts progression to chronic kidney disease. Kidney Int. 2011;79(12): 1361-9; Peerapornratana et al. Sepsis-Associated Acute Kidney Disease. Kidney Int Rep. 2020;5(6):839-50). an increased risk of cardiovascular complications, recurrent episodes of AKI, and long-term mortality (Hoste et al. 2015. Epidemiology of acute kidney injury in critically ill patients: the multinational AKI-EPI study. Intensive Care Medicine. 41(8): 1411-23; James et al. Long-term outcomes of acute kidney injury and strategies for improved care. Nature Reviews Nephrology. 2020; 16(4): 193-205).

[0090] The first international interdisciplinary consensus criteria for diagnosis of AKI were the RIFLE criteria (Bellomo et al. Acute renal failure — definition, outcome measures, animal models, fluid therapy and information technology needs: the Second International Consensus Conference of the Acute Dialysis Quality Initiative (ADQI) Group. Crit Care 2004; 8: R204-212). The acronym RIFLE stands for the increasing severity classes Risk, Injury, and Failure; and the two outcome classes, Loss and End-Stage Renal Disease (ESRD). The three severity grades are defined on the basis of the changes in SCr or urine output where the worst of each criterion is used. The two outcome criteria, Loss and ESRD, are defined by the duration of loss of kidney function.

[0091] The RIFLE criteria define 3 categories of impairment (risk, injury, and failure) and 2 categories of longterm renal outcomes (loss and end-stage renal disease (ESRD)). Whichever criterion shows the most impairment is used for classification. When baseline creatine (Cr) is unknown, a baseline glomerular filtration rate (GFR) between 75 and 100 mL / min is assumed, or the Modification of Diet in Renal Disease (MDRD) equation can be used to calculate an estimated baseline Cr.

[0092] 1. Risk: Cr j1of 1.5x baseline, GFR J, of 25%, or urine output (u / o) <0.5mL / kg / h for 6 h

[0093] 2. Injury: Cr j of 2x baseline, GFR J, of 50%, or urine output (u / o) <0.5mL / kg / h for 12 h

[0094] 3. Failure: Cr j of 3x baseline, GFR J, of 75 %, Cr >4.0, or urine output (u / o) <0.5mL / kg / h for 12 h

[0095] 4. Loss: Loss of kidney function for over 4 weeks

[0096] 5. ESRD: Loss of kidney function for over 3 months

[0097] It is widely accepted that GFR is the most useful overall index of kidney function in health and disease, and changes in SCr and urine output are surrogates for changes in GFR. In clinical practice, an abrupt decline in GFR is assessed from an increase in SCr or oliguria. Recognizing the limitations of the use of a decrease in kidney function for the early detection and accurate estimation of renal injury, there is a broad consensus that, while more sensitive and specific biomarkers are needed, changes in SCr and / or urine output form the basis of all diagnostic criteria for AKI.

[0098] In 2007, the AKI network (AKIN), a multi-disciplinary international group, proposed some small modifications to the RIFLE criteria (Mehta et al. Acute Kidney Injury Network: report of an initiative to improve outcomes in acute kidney injury. Crit Care 2007: 11: R31). The Risk, Injury, and Failure stages became stage 1, 2, and 3, and a >0.3 mg / dl rise in creatinine within 48 h was included in stage 1. GFR criteria were removed as markers of adult AKI. The Loss and End-stage categories of RIFLE were dropped as they were felt to be outcomes, not stages.

[0099] AKI according to KDIGO guidelines is defined as any of the following:

[0100] • Increase in SCr by > 0.3 mg / dl (> 26.5 pmol / 1) within 48 hours; or

[0101] • Increase in SCr to > 1.5 times baseline, which is known or presumed to have occurred within the prior 7 days, or

[0102] • Urine volume <0.5 ml / kg / h for 6 hours

[0103] The current definition of AKI was released in 2012 by the AKI work group of the Kidney Disease Improving Global Outcomes (KDIGO) organization. Definition and staging of AKI are based on the Risk, Injury, Failure; Loss, End-Stage Renal Disease (RIFLE) and Acute Kidney Injury Network (AKIN) criteria and the recent International Kidney Disease Improving Global Outcomes (KDIGOs) guideline merged RIFLE and AKIN and is based on an increase in sCr and / or a decrease in urine output as surrogate markers of the GFR (KDIGO. KDIGO clinical practice guideline for acute kidney injury. Kidney Int Suppl (2011). 2012:2(l):19-36). The term “acute kidney injury (AKI)” has been proposed to encompass the entire spectrum of the syndrome from minor changes in markers of renal function to requirement for renal replacement therapy (RRT).

[0104] Acute kidney failure is defined as a GFR <15 ml / min per 1.73 m2body surface area, or requirement for RRT, although it is recognized that RRT may be required earlier in the evolution of AKI.

[0105] In particular embodiments of the invention, acute kidney failure is defined as a GFR <15 ml / min per 1.73 m2body surface area, or requirement for RRT.

[0106] In particular embodiments of the invention, AKI is defined (according to KDIGO guidelines) as any of the following:

[0107] • Increase in SCr by > 0.3 mg / dl (> 26.5 pmol / 1) within 48 hours; or

[0108] • Increase in SCr to > 1.5 times baseline, which is known or presumed to have occurred within 7 days prior to the diagnosis, or

[0109] • Urine volume <0.5 ml / kg / h for 6 consecutive hours AKI according to KDIGO guidelines is staged for severity according to the following criteria (Table 1):

[0110] Table 1: The stages of AKI according to KDIGO guidelines.

[0111] Stage Serum creatinine Urine output

[0112] 1 1.5-1.9 times baseline < 0.5 ml / kg / h for

[0113] OR 6-12 hours

[0114] < 03 mg / dl ( 26.5 umol / l) increase

[0115] 2 2.0-2.9 times baseline <0.5 ml / kg / h for

[0116] / •r 12 hours

[0117] 3 3.0 times baseline < 03 ml / kg / h for

[0118] OR > 24 hours

[0119] Increase in serum creatinine to OR hours

[0120] Initiation of renal replacement therapy

[0121] OR, In patients < 18 years, decrease in eGFR to < 35 ml / min per 1.73 m2

[0122] The term “acute kidney injury” (AKI) according to the present invention in particular also encompasses acute kidney failure.

[0123] Glomerular filtration cannot be measured directly in humans; thus “true” GFR cannot be known with certainty. However, GFR can be assessed from clearance measurements (measured GFR [mGFR]) or serum levels of endogenous filtration markers (estimated GFR [eGFR]).

[0124] In particular embodiments of the invention, kidney function is determined by measuring the GFR, wherein more particularly GFR is equal to the total of the filtration rates of the functioning nephrons, and which is more particularly recorded in units of volume per time, e.g., millilitres per minute (mL / min). More particularly, GFR is determined by measuring clearance (measured GFR [mGFR]) or serum levels of endogenous filtration markers (estimated GFR [eGFR]).

[0125] The creatinine clearance rate (CCr or CrCl) is the volume of blood plasma that is cleared of creatinine per unit time and is a useful measure for approximating the GFR. Creatinine clearance exceeds GFR due to creatinine secretion, which can be blocked by cimetidine. Both GFR and CCr may be accurately calculated by comparative measurements of substances in the blood and urine or estimated by formulas using just a blood test result (eGFR and eCCr). The results of these tests are used to assess the excretory function of the kidneys. Estimated GFR (eGFR) is currently recommended by clinical practice guidelines and regulatory agencies for routine evaluation of GFR whereas measured GFR (mGFR) is recommended as a confirmatory test when more accurate assessment is required (Levey et al. 2020. Nat Rev Nephrol. 16 (1): 51-64).

[0126] There are several methods known in the art to determine eGFR. Estimated GFR may be calculated for example using CKD-EPI creatinine equation, CKD-EPI cystatin C equation or CKD-EPI creatinine cystatin C equation, the Modification of Diet in Renal Disease (MDRD) study equation, and Cockcroft- Gault equation (Santos and Martins 2015. World J Nephrol 4(3): 345-353).

[0127] In particular embodiments of the invention, creatinine clearance rate is measured to determine the GFR, wherein creatinine secretion is optionally blocked, e.g. by administration of cimetidine to the subject in which creatinine clearance rate is measured, wherein particularly GFR and / or CCr are calculated based on the results obtained by comparative measurement of substances in a blood sample and a urine sample obtained from the, or determined from a blood sample obtained from the subject. In more particular embodiments of the invention, estimated GFR is calculated using CKD-EPI creatinine equation, CKD- EPI cystatin C equation or CKD-EPI creatinine cystatin C equation, the Modification of Diet in Renal Disease (MDRD) study equation, or Cockcroft-Gault equation.

[0128] Several biomarkers have been proposed to aid in the diagnosis of AKI, e.g. proenkephalin 119-159 (also called penKid), cystatin C (CysC), KIM-1 (Kidney Injury Molecule-1), TIMP-2 (Tissue inhibitor of metalloproteinase), IGFBP-7 (Insulin-like growth factor-binding protein 7), blood urea nitrogen (BUN) and Neutrophil Gelatinase-Associated Lipocalin (NGAL).

[0129] In particular embodiments of the invention, the diagnosis of AKI is based on the determination of one or more biomarkers, e.g. biomarkers selected from the group comprising proenkephalin 119-159 (also called penKid), cystatin C (CysC), KIM-1 (Kidney Injury Molecule-1), TIMP-2 (Tissue inhibitor of metalloproteinase), IGFBP-7 (Insulin-like growth factor-binding protein 7), blood urea nitrogen (BUN) and Neutrophil Gelatinase-Associated Lipocalin (NGAL). These biomarkers and how AKI is diagnosed with them is known from the prior art.

[0130] Various clinical studies have shown that penKid strongly correlates with GFR over the full range (Beunders et al. Assessing GFR With Proenkephalin. Kidney Int Rep. 2023:8(11):2345-55). facilitates early detection of AKI (Hollinger et al. Proenkephalin A 119-159 (PenKid) Is an Early Biomarker of Septic Acute Kidney Injury: The Kidney in Sepsis and Septic Shock (Kid-SSS) Study. Kidney Int Rep. 2018:3(6): 1424-33). The performance of penKid in diagnosing AKI was summarized in a meta-analysis using data from patients with various critical conditions, such as sepsis, acute heart failure, and cardiac surgery incorporated 11 observational studies with 3969 patients with an incidence of AKI of 23.4% with the best optimal cutoff value of penKid for early detection of AKI being 57.3 pmol / L (Lin et al. Proenkephalin as a biomarker correlates with acute kidney injury: a systematic review with metaanalysis and trial sequential analysis. Crit Care. 2023;27(l):481 Moreover, the performance of penKid as an aid in the early diagnosis of AKI in a patient cohort with sepsis and septic shock has been evaluated recently (Schulte et al. 2024. Clinical performance of proenkephalin A 119-159 for the early diagnosis of acute kidney injury inpatients with sepsis or septic shock medRXiv preprint}. PenKid cutoff values of 54 pmol / L (>92% sensitivity) and 105 pmol / L (>92% specificity) were derived to establish actionable interpretation bands for diagnostic rule-in and rule-out.

[0131] Therefore, in particular embodiments of the invention, AKI is diagnosed via GFR, sCr, sCr clearance, urine output, proenkephalin 119-159, cystatin C (CysC), KIM-1, TIMP-2, IGFBP-7, blood urea nitrogen (BUN) and / or Neutrophil Gelatinase-Associated Lipocalin (NGAL).

[0132] In particular embodiments of the invention, improvement of kidney function is defined as any improvement of one or more kidney parameters, for example: improvement in categorization or staging of AKI (e.g. RIFLE, AKIN, KDIGO), increase in glomerular filtration rate (GFR), decrease of serum creatinine (sCr), increase in serum creatinine clearance, increase in urine output, and / or decrease of AKI biomarkers (e.g. Pro-Enkephalin 119-159, CysC, KIM-1, TIMP-2, IGFBP-7, BUN and / or NGAL). This may in particular be determined by comparing the results of two or more measurements of said kidney parameters in the same subject.

[0133] In specific embodiments of the invention improvement of kidney function is defined as an improvement of GFR, wherein said GFR increase is at least 1%, more preferred at least 5%, even more preferred at least 10%, even more preferred at least 15%, even more preferred at least 25%, even more preferred at least 40%, most preferred said GFR increase is at least 50%.

[0134] In specific embodiments of the invention improvement of kidney function is defined as an improvement of sCr, wherein said sCr decrease is at least 1%, more preferred at least 5%, even more preferred at least 10%, even more preferred at least 15%, even more preferred at least 25%, even more preferred at least 40%, most preferred said sCr decrease is at least 50%.

[0135] In specific embodiments of the invention improvement of kidney function is defined as an improvement of sCr clearance, wherein said sCr clearance increase is at least 1%, more preferred at least 5%, even more preferred at least 10%, even more preferred at least 15%, even more preferred at least 25%, even more preferred at least 40%, most preferred said sCr clearance increase is at least 50%. In specific embodiments of the invention improvement of kidney function is defined as an improvement of urine output, wherein said increase in urine output is at least 1%, more preferred at least 5%, even more preferred at least 10%, even more preferred at least 15%, even more preferred at least 25%, even more preferred at least 40%, most preferred said urine output increase is at least 50%.

[0136] In specific embodiments of the invention improvement of kidney function is defined as an improvement of AKI biomarker, wherein said decrease in AKI biomarkers (e.g. Pro-Enkephalin 119-159, CysC, KIM- 1, TIMP-2, IGFBP-7, BUN and / or NGAL) is at least 1%, more preferred at least 5%, even more preferred at least 10%, even more preferred at least 15%, even more preferred at least 25%, even more preferred at least 40%, most preferred said decrease in AKI biomarker is at least 50%.

[0137] In specific embodiments of the invention, improvement of kidney function is defined as an improvement of AKI biomarker, wherein said AKI biomarker (e.g. Pro-Enkephalin 119-159, CysC, KIM-1, TIMP-2, IGFBP-7, BUN and / or NGAL) is decreased below a threshold.

[0138] In specific embodiments of the invention improvement of kidney function is defined as an improvement of the AKI biomarker proenkephalin 119-159, wherein said threshold of proenkephalin 119-159 is in the range between 50 and 120 pmol / L, preferably in the range between 50 and 100 pmol / 1, most preferred in the range between 50 and 80 pmol / L.

[0139] In particular embodiments of the invention, stabilization of kidney function means no further worsening of kidney function after therapy of the respective patient. Worsening of kidney function is defined as any worsening of one or more kidney parameters, for example: worsening in categorization or staging of AKI (e.g. RIFLE, AKIN, KDIGO), decrease in glomerular filtration rate (GFR), increase of serum creatinine (sCr), decrease in serum creatinine clearance, decrease in urine output, and / or increase of AKI biomarkers (e.g. Pro-Enkephalin 119-159, CysC, KIM-1, TIMP-2, IGFBP-7, BUN and / or NGAL). This may in particular be determined by comparing the results of two or more measurements of said kidney parameters in the same subject.

[0140] In particular embodiments of the invention, an improvement and / or stabilization of kidney function will occur within 7 days or less, preferably within 5 days or less, more preferred within 48 hours or less, even more preferred within 24 hours or less, most preferred within 12 hours or less upon starting therapy with an anti-DPP3 antibody or fragment thereof binding to DPP3, wherein in particular the start of therapy is after the first dose has been completed, e.g., after infusion of the anti-DPP3 antibody has been completed for the first dose.

[0141] In one embodiment of the invention said extreme-critically ill patient is characterized by a level of DPP3 in a sample of a bodily fluid of said patient above a threshold. In one embodiment, said threshold is a level of DPP3 in a sample of a bodily fluid of said patient, wherein the DPP3 concentration is between 20 and 120 ng / mL, more preferred between 25 and 100 ng / mL, even more preferred between 30 and 80 ng / mL, even more preferred between 35 and 60 ng / mL, even more preferred between 40 and 50 ng / ml most preferred said threshold is 40 ng / mL.

[0142] In another embodiment said threshold is a level of DPP3 in a sample of bodily fluid of said patient, wherein the DPP3 activity is between 1.76 and 9.68 pmol / (min*L), more preferred between 2.15 and 8.09 pmol / (min*L), even more preferred between 2.55 and 6.51 pmol / (min*L), even more preferred between 2.94 and 4.93 pmol / (min*L), even more preferred between 3.34 and 4.13 pmol / (min*L), most preferred said threshold is 3.34 pmol / (min*L).

[0143] In a specific embodiment said sample of bodily fluid is taken from said patient before said treatment with a DPP3 antibody is initiated.

[0144] As used herein, “DPP3 antibody” and “anti-DPP3 antibody” are used interchangeably.

[0145] In one aspect of the invention the threshold from said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of DPP 3 in said sample.

[0146] DPP3 activity and cDPP3 activity are generically described herein as DPP3 activity. In general, if DPP3 activity is measured in a bodily fluid, such as the blood, this means that cDPP3 activity is being measured.

[0147] DPP3 concentration and cDPP3 concentration are generically described herein as DPP3 concentration. In general, if DPP3 concentration is measured in a bodily fluid, such as the blood, this means that cDPP3 concentration is being measured.

[0148] DPP3 activity can be measured by detection of cleavage products of DPP3 specific substrates. Known peptide hormone substrates include Leu-enkephalin, Met-enkephalin, endomorphin 1 and 2, valorphin, P-casomorphin, dynorphin, proctolin, ACTH (Adrenocorticotropic hormone) and MSH (melanocytestimulating hormone (M. Abramic et al. 2000) (Barsun et al. 2007) (Dhanda, Singh, and Singh 2008). The cleavage of mentioned peptide hormones as well as other untagged oligopeptides (e.g., Ala-Ala- Ala-Ala, (Dhanda, Singh, and Singh 2008)) can be monitored by detection of the respective cleavage products. Detection methods include, but are not limited to, HPLC analysis (e.g., (Lee and Snyder 1982), mass spectrometry (e.g., (M. Abramic et al. 2000), Hl-NMR analysis (e.g., (Vandenberg, King, and Kuchel 1985), capillary zone electrophoresis (CE; e.g., (Barsun et al. 2007)), thin layer chromatography (e.g., (Dhanda, Singh, and Singh 2008)) or reversed phase chromatography (e.g., (Mazzocco et al. 2006)).

[0149] Detection of fluorescence due to hydrolysis of Anorogenic substrates by DPP3 is a standard procedure to monitor DPP3 activity. Those substrates are specific di- or tripeptides (Arg-Arg, Ala-Ala, Ala-Arg, Ala-Phe, Asp-Arg, Gly-Ala, Gly-Arg, Gly-Phe, Leu-Ala, Leu-Gly, Lys-Ala, Phe-Arg, Suc-Ala-Ala-Phe) coupled to a Auorophore. Fluorophores include but are not limited to P-naphtylamide (2-naphtylamide, PNA, 2NA), 4-methoxy-P-naphtylamide (4-methoxy-2-naphtylamide) and 7-amido-4-methylcoumarin (AMC, MCA; (M. Abramic et al. 2000) (Ohkubo et al. 1999).

[0150] Cleavage of these Anorogenic substrates leads to the release of Auorescent P-naphtylamine or 7-amino- 4-methylcoumarin respectively. In a liquid phase assay or an ECA substrate and DPP3 are incubated in for example a 96 well plate format and Auorescence is measured using a Auorescence detector (Ellis and Nuenke 1967).

[0151] Additionally, DPP3 carrying samples can be immobilized and divided on a gel by electrophoresis, gels stained with Anorogenic substrate (e.g., Arg-Arg-pNA) and Fast Garnet GBC and Auorescent protein bands detected by a Auorescence reader (Ohkubo et al. 1999). The same peptides (Arg-Arg, Ala-Ala, Ala-Arg, Ala-Phe, Asp-Arg, Gly-Ala, Gly-Arg, Gly-Phe, Leu-Ala, Leu-Gly, Lys-Ala, Phe-Arg, Suc-Ala- Ala-Phe) can be coupled to chromophores, such as p-nitroanilide diacetate. Detection of color change due to hydrolysis of chromogenic substrates can be used to monitor DPP3 activity.

[0152] Another option for the detection of DPP3 activity is a Protease-Gio™ Assay (commercially available at Promega). In this embodiment of said method DPP3 specific di- or tripeptides (Arg-Arg, Ala-Ala, Ala- Arg, Ala-Phe, Asp-Arg, Gly-Ala, Gly-Arg, Gly-Phe, Leu-Ala, Leu-Gly, Lys-Ala, Phe-Arg, Suc-Ala- Ala-Phe) are coupled to aminoluciferin. Upon cleavage by DPP3, aminoluciferin is released and serves as a substrate for a coupled luciferase reaction that emits detectable luminescence.

[0153] In a preferred embodiment DPP3 activity is measured by addition of the Anorogenic substrate Arg -Arg - PNA and monitoring Auorescence in real time.

[0154] In another embodiment of the invention, the level of DPP3 is determined by a method comprising contacting said sample of bodily Auid with a capture binder that binds specifically to DPP3. In another preferred embodiment of the invention, said capture binder for determining the level of DPP3 may be selected from the group of antibody, antibody fragment or non-IgG scaffold.

[0155] In a specific embodiment of the invention, said capture binder for determining the level of DPP3 is an antibody.

[0156] Another specific embodiment of the invention comprises the use of a capture-binder that binds specifically to full-length DPP3.

[0157] In another preferred embodiment of the invention said capture-binder is immobilized on a solid phase. The test sample may be passed over the immobile binder, and DPP3, if present in the sample, binds to the binder and is itself immobilized for detection.

[0158] In one embodiment, a substrate may then be added, and the reaction product may be detected to indicate the presence or amount of DPP3 protein or activity in the test sample.

[0159] For the purposes of the present description, the term "solid phase" may be used to include any material or vessel in which or on which the assay may be performed and includes, but is not limited to porous materials, nonporous materials, test tubes, wells, slides, agarose resins (e.g., Sepharose from GE Healthcare Life Sciences), magnetic particles (e.g., Dynabeads™ or Pierce™ magnetic beads from Thermo Fisher Scientific), etc.

[0160] In one embodiment of the invention the method for determining DPP3 activity in a bodily fluid sample of said subject comprises the steps:

[0161] • contacting said sample with a capture-binder that binds specifically to full-length DPP3,

[0162] • separating DPP3 bound to said capture binder,

[0163] • adding substrate of DPP3 to said separated DPP3,

[0164] • quantifying of said DPP 3 activity by measuring and quantifying the conversion of a substrate of DPP3.

[0165] In another embodiment of the invention said separation step is a washing step that removes ingredients of the sample that are not bound to said capture-binder from the captured DPP3.

[0166] In another embodiment of the invention the DPP3 substrate conversion is detected by a method selected from the group comprising: fluorescence of fluorogenic substrates (e.g. Arg-Arg-PNA, Arg-Arg-AMC), color change of chromogenic substrates, luminescence of substrates coupled to aminoluciferin, mass spectrometry, HPLC / FPLC (reversed phase chromatography, size exclusion chromatography), thin layer chromatography, capillary zone electrophoresis, gel electrophoresis followed by activity staining (immobilized, active DPP3) or western blot (cleavage products).

[0167] In another embodiment of the invention said substrate may be selected from the group comprising: angiotensin II, III and IV, Leu-enkephalin, Met-enkephalin, endomorphin 1 and 2, valorphin, P- casomorphin, dynorphin, proctolin, ACTH and MSH, or di-peptides coupled to a fluorophore, a chromophore or aminoluciferin wherein the di-peptide is Arg -Arg.

[0168] In another specific embodiment of the invention said substrate may be selected from the group comprising: A di -peptide coupled to a fluorophore, a chromophore or aminoluciferin wherein the dipeptide is Arg -Arg.

[0169] In a specific embodiment, said capture-binder exhibits a binding affinity to DPP3 of at least 107M1, preferred 108M1, more preferred affinity is greater than 109M1, most preferred greater than IO10M1. A person skilled in the art knows that it may be considered to compensate lower affinity by applying a higher dose of compounds and this measure would not lead out-of-the-scope of the invention.

[0170] In one embodiment, said capture-binder is an antibody binding to DPP3.

[0171] To determine the affinity of the antibodies to DPP3, in one embodiment the kinetics of binding of DPP3 to an immobilized antibody are determined by means of label-free surface plasmon resonance using a Biacore 2000 system (GE Healthcare Europe GmbH, Freiburg, Germany). Reversible immobilization of the antibodies was performed using an anti-mouse Fc antibody covalently coupled in high density to a CM5 sensor surface according to the manufacturer's instructions (mouse antibody capture kit; GE Healthcare), (Lorenz etal. 2011. Antimicrob Agents Chemother. 55 (1): 165 173}.

[0172] Alternatively, the DPP3 bound to a capture binder on a solid phase is detected with a second capture binder specifically binding to DPP3.

[0173] In one embodiment such assay for determining the level of DPP3 is a sandwich immunoassay using any kind of detection technology including but not restricted to enzyme label, chemiluminescence label, electrochemiluminescence label, preferably a fully automated assay. In one embodiment of the diagnostic method such an assay is an enzyme labeled sandwich assay. Examples of automated or fully automated assay comprise assays that may be used for one of the following systems: Roche Elecsys®, Abbott Architect®, Siemens Advia Centauer®, Siemens Immulite®, Brahms Kryptor®, Biomerieux Vidas®, Alere Triage®, Boditech AFIAS®, Ortho Vidas®, Diasorin LIASION®, Beckman Dxl®, Lumira Dx®, MeMed Key®, Werfen BioFlash®, BioRad BioPlex®. A variety of immunoassays are known and may be used for the assays and methods of the present invention. In one embodiment of the present invention, the immunoassays is selected from the group comprising mass spectrometry (MS), luminescence immunoassay (LIA), immunoluminometric assay (ILMA), radioimmunoassays ("RIA"), homogeneous enzyme-multiplied immunoassays ("EMIT"), enzyme linked immunoadsorbent assays ("ELISA"), apoenzyme reactivation immunoassay ("ARIS"), chemiluminescence- (“CLIA”), electrochemiluminescence- (“ECLIA”) and fluorescenceimmunoassays, luminescence-based bead arrays, magnetic beads based arrays, protein microarray assays, rapid test formats such as for instance dipstick immunoassays, immuno-chromatographic strip tests, rare cryptate assay and automated systems / analyzers.

[0174] In one embodiment of the invention the assay or method may be a so-called POC (point-of-care) -test that is a test technology, which allows performing the test within less than 1 hour near the patient without the requirement of a fully automated assay system. One example for this technology is the immunochromatographic test technology, e.g., a microfluidic device.

[0175] In a specific embodiment at least one of said two binders is labeled in said sandwich immunoassay in order to be detected.

[0176] In another preferred embodiment said label is selected from the group comprising chemiluminescent label, enzyme label, fluorescence label, radioiodine label.

[0177] The assays can be homogenous or heterogeneous assays, competitive and non-competitive assays. In one embodiment, the assay is in the form of a sandwich assay, which is a non-competitive immunoassay, wherein the molecule to be detected and / or quantified is bound to a first antibody and to a second antibody. The first antibody may be bound to a solid phase, e.g. a bead, a surface of a well or other container, a chip or a strip, and the second antibody is an antibody which is labeled, e.g. with a dye, with a radioisotope, or a reactive or catalytically active moiety. The amount of labeled antibody bound to the analyte is then measured by an appropriate method. The general composition and procedures involved with “sandwich assays” are well-established and known to the skilled person (The Immunoassay Handbook, Ed. David Wild, Elsevier LTD, Oxford; 3rd ed. (May 2005), ISBN-13: 978-0080445267; Hultschig C etal., Curr Opin Chem Biol. 2006 Feb;10(l):4-10. PMID: 16376134).

[0178] In another embodiment the assay comprises two capture molecules, preferably antibodies which are both present as dispersions in a liquid reaction mixture, wherein a first labelling component is attached to the first capture molecule, wherein said first labelling component is part of a labelling system based on fluorescence- or chemiluminescence-quenching or amplification, and a second labelling component of said marking system is attached to the second capture molecule, so that upon binding of both capture molecules to the analyte a measurable signal is generated that allows for the detection of the formed sandwich complexes in the solution comprising the sample.

[0179] In another embodiment, said labeling system comprises rare earth cryptates or rare earth chelates in combination with fluorescence dye or chemiluminescence dye, in particular a dye of the cyanine type.

[0180] In an embodiment of the present invention, fluorescence-based assays comprise the use of dyes, are selected from the group comprising FAM (5 -or 6-carboxyfluorescein), VIC, NED, Fluorescein, Fluoresceinisothiocyanate (FITC), IRD-700 / 800, Cyanine dyes, such as CY3, CY5, CY3.5, CY5.5, Cy7, Xanthen, 6-Carboxy-2’,4’,7’,4,7-hexachlorofluorescein (HEX), TET, 6-Carboxy-4’,5’-dichloro-2’,7’- dimethodyfluorescein (JOE), N,N,N’,N’-Tetramethyl-6-carboxyrhodamine (TAMRA), 6-Carboxy-X- rhodamine (ROX), 5-Carboxyrhodamine-6G (R6G5), 6-carboxyrhodamine-6G (RG6), Rhodamine, Rhodamine Green, Rhodamine Red, Rhodamine 110, BODIPY dyes, such as BODIPY TMR, Oregon Green, Coumarines such as Umbelliferone, Benzimides, such as Hoechst 33258; Phenanthridines, such as Texas Red, Yakima Yellow, Alexa Fluor, PET, Ethidiumbromide, Acridinium dyes, Carbazol dyes, Phenoxazine dyes, Porphyrine dyes, Polymethin dyes, and the like.

[0181] In the context of the present invention, chemiluminescence based assays comprise the use of dyes, based on the physical principles described for chemiluminescent materials in (Kirk-Othmer, Encyclopedia of chemical technology, 4th ed., executive editor, J. I. Kroschwitz; editor, M. Howe-Grant, John Wiley & Sons, 1993, vol,15, p. 518-562, incorporated herein by reference, including citations on pages 551-562}, Preferred chemiluminescent dyes are acridiniumesters.

[0182] As mentioned herein, an “assay” or “diagnostic assay” can be of any type applied in the field of diagnostics.

[0183] Such an assay may be based on the binding of an analyte to be detected to one or more capture probes with a certain affinity. Concerning the interaction between capture molecules and target molecules or molecules of interest, in one embodiment the affinity constant is preferably greater than 108M1.

[0184] In a specific embodiment an assay is used for determining the level of DPP3, wherein the assay sensitivity of said assay is able to quantify the DPP3 of at least a portion of healthy subjects and is < 20 ng / ml, preferably < 30 ng / ml and more preferably < 40 ng / ml. In a specific embodiment an assay is used for determining the level of DPP3, wherein the assay sensitivity of said assay is able to quantify the DPP3 ofat least a portion of healthy subjects and is < the 1.4-fold of the median of the level ofDPP3 in a healthy population, preferably < the 2.1 -fold of the median of the level of DPP3 in a healthy population and more preferably < the 2.8 -fold of the median of the level of DPP3 in a healthy population. A bodily fluid according to the present invention is in one particular embodiment a blood sample. A blood sample is in particular selected from the group comprising whole blood, serum and plasma. In a specific embodiment of the method said sample is selected from the group comprising human citrate plasma, heparin plasma, and EDTA plasma.

[0185] In another specific embodiment of the present invention said level of DPP3 is determined in different samples taken from said patient at different time-points.

[0186] In another specific embodiment of the present invention the difference between said level of DPP3 in different samples taken from said patient at different time-points is determined. The difference may be determined as absolute or relative difference.

[0187] In a specific embodiment of the present invention said level of DPP3 is determined at least twice.

[0188] The level of DPP3 as DPP3 concentration and / or DPP3 activity in a sample of bodily fluid of said subject may in one embodiment of the present invention be determined for example by one of the following methods:

[0189] 1. Luminescence immunoassay for the quantification of DPP3 protein concentrations (LIA) (Rehfeld et al., 2019).

[0190] 2. Enzyme capture activity assay for the quantification ofDPP3 activity (ECA) (Rehfeld et al., 2019).

[0191] 3. Liquid-phase assay for the quantification of DPP3 activity (LAA) (modified from Jones et al., Analytical Biochemistry, 1982}

[0192] 4. Liquid-phase assay for the quantification of DPP3 activity (LAA) as described in Example 3.2.

[0193] 5. Point-of-care assay forthe quantification ofDPP3 protein concentration (IB10 sphingotest® DPP3) as described in example 3.1.

[0194] Threshold values mentioned above might be different in other assays, if these have been calibrated differently from the assay systems used in the present invention. Therefore, the mentioned threshold values above shall apply for such differently calibrated assays accordingly, taking into account the differences in calibration. One possibility of quantifying the difference in calibration is a method comparison analysis (correlation) of the assay in question with the respective biomarker assay used in the present invention by measuring the respective biomarker (e.g., DPP3) in samples using both methods. Assuming a linear correlation between the assays, another possibility is to determine with the assay in question, given this test has sufficient analytical sensitivity, the median biomarker level of a representative normal population, compare results with the median biomarker levels as described in the literature and recalculate the calibration based on the difference obtained by this comparison, e.g. by using a factor. Hence, the x-fold of e.g. the mean or median (or a specific percentile) of the level of DPP3 (either concentration or activity of DPP3) in a healthy population can be used as a threshold level using a differently calibrated assay.

[0195] With the calibration used in the present invention, samples from 5,400 normal (healthy) subjects (swedish single-center prospective population-based Study (MPP-RES)) have been measured: median (interquartile range) plasma DPP3 concentration was 14.5 ng / ml (11.3 ng / ml - 19 ng / ml). The corresponding DPP3 activity was 1.32 (1.07-1.68) pmol / (min*L).

[0196] In one embodiment of the present invention said threshold of DPP3 concentration is between 20 and 120 ng / mL, more preferred between 25 and 100 ng / mL, even more preferred between 30 and 80 ng / mL, even more preferred between 35 and 60 ng / mL, even more preferred between 40 and 50 ng / ml, most preferred said threshold is 40 ng / mL.

[0197] In one embodiment of the present invention said threshold of DPP3 activity is between 1.76 and 9.68 pmol / (min*L), more preferred between 2.15 and 8.09 pmol / (min*L), even more preferred between 2.55 and 6.51 pmol / (min*L), even more preferred between 2.94 and 4.93 pmol / (min*L), even more preferred between 3.34 and 4.13 pmol / (min*L), most preferred said threshold is 3.34 pmol / (min*L).

[0198] In another embodiment of the invention said threshold is an x-fold of the median level of DPP3 concentration in a healthy population. In a specific embodiment of the invention the threshold of DPP3 concentration is in the range between the 1.4-fold and 8.3-fold, more preferred in the range between the 1.7-fold and 6.9-fold, more preferred in the range between 2.1-fold and 5.5-fold, even more preferred in the range between 2.4 and 4.1, even more preferred in the range between 2.8 and 3.4, most preferred said threshold is the 2.8-fold of the median of the DPP3 concentration in a healthy population.

[0199] In one embodiment of the present invention said threshold of DPP3 concentration is 120 ng / ml or less, more preferred 100 ng / mL or less, even more preferred 80 ng / mL or less, even more preferred 60 ng / ml or less, even more preferred 50 ng / ml or less, most preferred said threshold is 40 ng / mL.

[0200] In a specific embodiment of the invention the threshold of DPP3 concentration is the 8.3-fold or less, more preferred the 6.9-fold or less, even more preferred the 5.5-fold or less, even more preferred the 4.1-fold or less, even more preferred the 3.4-fold or less, most preferred the 2.8-fold of the median of the DPP3 concentration in a healthy population.

[0201] In one embodiment of the present invention said threshold of DPP3 activity is 9.68 pmol / (min*L) or less, more preferred 8.09 pmol / (min*L) or less, even more preferred 6.51 pmol / (min*L) or less, even more preferred 4.93 pmol / (min*L) or less, even more preferred 4.13 pmol / (min*L) or less, most preferred said threshold is 3.34 pmol / (min*L).

[0202] In a specific embodiment of the invention the threshold of DPP3 activity is the 8.3-fold or less, more preferred the 6.9-fold or less, even more preferred the 5.5-fold or less, even more preferred the 4.1-fold or less, even more preferred the 3.4-fold or less, most preferred the 2.8-fold of the median of the DPP3 activity in a healthy population.

[0203] The DPP3 activity was defined and measured as described in example 3.2.

[0204] The threshold level is a level, which allows for allocating the patient into a group of patients who have been diagnosed and / or are having an increased risk of an adverse event (e.g. mortality), or into a group of patients who have not been diagnosed and / or have a decreased risk of an adverse event, or into a severity group.

[0205] Thus, the threshold level shall allow for differentiating between a patient who is diagnosed and / or is having an increased risk of an adverse event and a patient who is not diagnosed and / or is having a decreased risk of an adverse event.

[0206] It is known in the art how threshold levels of an analyte can be determined. Threshold levels are predetermined levels of an analyte to which actually measured levels from a patient are compared. Depending on whether the patient’s measured analyte level is below or above the predetermined threshold level, the patient is categorized as having or not having a certain diagnosis or increased risk of an adverse event, depending on the specific analyte and clinical question to be addressed. Threshold levels are set to meet routine clinical requirements. These requirements can vary. If for instance it is clinically more relevant to achieve a high diagnostic sensitivity at the expense of a relatively low diagnostic specificity, the threshold level might be chosen differently than if just the opposite, e.g. high diagnostic specificity at the expense of a relatively low diagnostic sensitivity, shall be achieved. It may for example be that sensitivity or specificity, respectively, has to be set to certain limits, e.g., 80%, 90%, 95% or 98%, respectively.

[0207] The sensitivity and specificity of a diagnostic and / or prognostic test depends on more than just the analytical "quality" of the test, they also depend on the definition of what constitutes an abnormal result. In practice, Receiver Operating Characteristic curves (ROC curves), are typically calculated by plotting the value of a variable versus its relative frequency in "reference group" (i.e. patients who do not have the disease) and "disease" populations (i.e. patients who have the disease). For any particular biomarker, a distribution of biomarker levels for patients with and without disease will likely overlap. Under such conditions, a test does not absolutely distinguish patients with and without disease (e.g., shock) with 100% accuracy, and the area of overlap indicates where the test cannot distinguish normal from disease. A threshold is selected, above which (or below which, depending on how a biomarker changes with the disease) the test is considered to be abnormal and below which the test is considered to be normal. The area under the ROC curve is a measure of the probability that the perceived measurement will allow correct identification of a condition. ROC curves can be used even when test results do not necessarily give an accurate number. As long as one can rank results, one can create a ROC curve. For example, results of a test on "disease" samples might be ranked according to degree (e.g. Mow, 2=normal, and 3=high). This ranking can be correlated to results in the "reference" group, and a ROC curve created. These methods are well known in the art (See, e.g., Hanley et al.1982. Radiology 143: 29-36). Preferably, ROC curves result in an Area under the ROC curve (AUC) of greater than about 0.5, more preferably greater than about 0.7, still more preferably greater than about 0.8, even more preferably greater than about 0.85, and most preferably greater than about 0.9. The term "about" in this context refers to + / - 5% of a given measurement.

[0208] The horizontal axis of the ROC curve represents (1 -specificity), which increases with the rate of false positives. The vertical axis of the curve represents sensitivity, which increases with the rate of true positives.

[0209] Thus, for a particular cut-off threshold selected, the value of (1 -specificity) (in words, “one minus specificity”) may be determined, and a corresponding sensitivity may be obtained. The area under the ROC curve is a measure of the probability that the measured biomarker level will allow correct identification of a disease or condition. Thus, the area under the ROC curve can be used to determine the effectiveness of the test.

[0210] In addition to the normal range, other methods may be used to determine thresholds for a specific indication, depending on the intended use and application / clinical setting. Such methods include e.g. the Youden optimum, thresholds that maximise overall accuracy, the odds ratio, or the positive or negative predictive value. In some situations, thresholds achieving a pre-specified level of sensitivity or specificity (e.g. 80%, 90%, 95% or 99%) can be appropriate for the clinical application. The choice of methods depends on the clinical application, which weights the costs of false positive and false negative results based on the test result consequences for the patient and the health care system, as well as the clinical need. Finally, multiple approaches may be combined to define a consensus threshold.

[0211] A “reference group” may be a healthy population, e.g., with no signs and symptoms of a disease. A reference group may consist of more than one reference subjects, i.e. healthy subjects. Preferred threshold values are for instance the 90th, 95th, or 99th percentile of a reference group (e.g. healthy population). By using a higher percentile than the 75th percentile, one reduces the number of false positive subjects identified, but one might miss to identify subjects, who are at moderate, albeit still increased risk. Thus, one might adopt the threshold value depending on whether it is considered more appropriate to identify most of the subjects at risk at the expense of also identifying "false positives", or whether it is considered more appropriate to identify mainly the subjects at high risk at the expense of missing several subjects at moderate risk. For example, the 75thpercentile, more preferred the 90thpercentile, even more preferred a 95thpercentile, most preferred the 99thpercentile values can be used for the upper limits of the normal range.

[0212] Biomarkers can also be used for the prediction of a future event or risk (e.g. prediction of mortality risk). For time-to-event data threshold levels can be obtained for instance from a Kaplan-Meier analysis, where the occurrence of a disease is correlated with e.g. the tertiles, quartiles, quintiles of the biomarkers in the population.

[0213] There are also equivalent methods available to the ROC methods described before, based on i.e. timedependent ROC analysis or generalizations of the area under the ROC curve (C index).

[0214] In one embodiment of the invention the successful treatment of a patient with shock may result in a reduced requirement of, e.g. <0.2pg / kg / body weight, or halving of initial norepinephrine dosing to keep the mean arterial pressure (MAP) above the target of 65 mmHg. Such reduced requirement can be coined “reversal of shock”.

[0215] In one embodiment of the invention an anti-DPP3 antibody or an anti-DPP3 antibody fragment is used for the treatment of shock in a patient having shock, wherein said patient is an extreme-critically ill patient.

[0216] In one embodiment of the invention said anti-DPP3 antibody is a humanized monoclonal IgGl antibody with a heavy chain consisting of or comprising the sequence SEQ ID NO.: 12 and with a light chain consisting of the sequence SEQ ID NO.: 13.

[0217] In one embodiment of the invention said anti-DPP3 antibody is a humanized monoclonal IgGl antibody with a heavy chain consisting of or comprising the sequence SEQ ID NO.: 34 and with a light chain consisting of the sequence SEQ ID NO.: 35.

[0218] Said anti-DPP3 antibody specifically binds circulating DPP3, targets and modulates DPP3 activity, an essential regulator of cardiovascular function. Its mode of action is relevant in acute diseases that are associated with massive cell death and uncontrolled release of intracellular DPP3 into the bloodstream. Translocated DPP3 remains active in the circulation where it cleaves bioactive peptides in an uncontrolled manner. In one embodiment of the present invention, the anti-DPP3 antibody is a humanized anti-DPP3 antibody. The anti-DPP3 antibody and fragments thereof according to the present invention, including a humanized anti-DPP3 antibody, is able to block circulating DPP3, inhibiting bioactive peptide degradation in the bloodstream. This blockade results in stabilization of cardiovascular and renal function and reduction of short-term mortality. Preclinical studies of anti-DPP3 antibodies in animal models of cardiovascular failure showed impressive and instant efficacy. As an example, injection of a humanized anti-DPP3 antibody in mice with shock-induced cardiovascular failure led to an instant normalization of the shortening fraction (Deniau et al. 2020). In several preclinical cardiovascular failure models, a humanized anti-DPP3 antibody has shown to improve all clinically relevant endpoints in vivo (Garcia et al. 2024). In one embodiment, the anti-DPP3 antibody or a fragment thereof according to the present invention normalizes ejection fraction and kidney function and reduces mortality.

[0219] Throughout the specification the “antibodies”, or “antibody fragments”, or “scaffolds” in accordance with the invention are capable to bind DPP3, and thus are directed against DPP3, and thus can be referred to as “anti-DPP3 antibodies”, “anti-DPP3 antibody fragments”, or “anti-DPP3 scaffolds”.

[0220] The term “antibody” generally comprises monoclonal and polyclonal antibodies and binding fragments thereof, in particular Fc-fragments as well as so called “single-chain-antibodies” (Bird et al. 1988), chimeric, humanized, in particular CDR-grafted antibodies, and dia or tetrabodies (Holliger, Prospero, and Winter 1993). Thus, in one embodiment of the present invention, the anti-DPP3 antibody has a format selected from the group comprising Fc-fragments single -chain-antibodies, chimeric antibodies, humanized, CDR-grafted antibodies, dia or tetrabodies, In a further embodiment, the anti-DPP3 antibody according to the present invention is a monoclonal or polyclonal antibody.

[0221] Also comprised are immunoglobulin-like proteins that are selected through techniques including, for example, phage display to specifically bind to the molecule of interest contained in a sample. In this context the term “specific binding” refers to antibodies raised against the molecule of interest or a fragment thereof.

[0222] An antibody is in particular considered to be specific, if its affinity towards the molecule of interest or the aforementioned fragment thereof is at least preferably 50-fold higher, more preferably 100-fold higher, most preferably at least 1000-fold higher than towards other molecules comprised in a sample containing the molecule of interest. It is well known in the art how to make antibodies and to select antibodies with a given specificity. In one embodiment of the invention the anti-DPP3 antibody or anti-DPP3 antibody fragment or anti- DPP3 non-Ig scaffold is monospecific.

[0223] Monospecific anti-DPP3 antibody or monospecific anti-DPP3 antibody fragment or monospecific anti- DPP3 non-Ig scaffold means that said antibody or antibody fragment or non-Ig scaffold binds to one specific region encompassing at least 5 amino acids within the target DPP3 (SEQ ID No. 1). Monospecific anti-DPP3 antibody or monospecific anti-DPP3 antibody fragment or monospecific anti- DPP3 non-Ig scaffold are anti-DPP3 antibodies or anti-DPP3 antibody fragments or anti-DPP3 non-Ig scaffolds that all have affinity for the same antigen. Monoclonal antibodies are monospecific, but monospecific antibodies may also be produced by other means than producing them from a common germ cell.

[0224] An antibody or fragment according to the present invention is a protein including one or more polypeptides substantially encoded by immunoglobulin genes that specifically binds an antigen. The recognized immunoglobulin genes include the kappa, lambda, alpha (IgA), gamma (IgGl, IgG2, IgG3, IgG4), delta (IgD), epsilon (IgE), and mu (IgM) constant region genes, as well as the myriad immunoglobulin variable region genes. Full-length immunoglobulin light chains are generally about 25 kDa or 214 amino acids in length.

[0225] Full-length immunoglobulin heavy chains are generally about 50 kDa or 446 amino acid in length. Light chains are encoded by a variable region gene at the NFE-terminus (about 110 amino acids in length) and a kappa or lambda constant region gene at the COOH-terminus. Heavy chains are similarly encoded by a variable region gene (about 116 amino acids in length) and one of the other constant region genes.

[0226] The basic structural unit of an antibody is generally a tetramer that consists of two identical pairs of immunoglobulin chains, each pair having one light and one heavy chain. In each pair, the light and heavy chain variable regions bind to an antigen, and the constant regions mediate effector functions. Immunoglobulins also exist in a variety of other forms including, for example, Fv, Fab, and (Fab')2, as well as bifimctional hybrid antibodies and single chains (c.v... I.anzavecchia et al. 1987. Eur. J. Immunol. 17: 105; Huston et al. 1988. Proc. Natl. Acad. Sci. U.S.A., 85: 5879-5883; Bird et al. 1988. Science 242: 423-426; Hood et al. 1984, Immunology, Benjamin, N.Y., 2nd ed. ; Hunkapiller and Hood 1986. Nature 323: 15-16}. An immunoglobulin light or heavy chain variable region includes a framework region interrupted by three hypervariable regions, also called complementarity determining regions (CDRs) (see, Sequences of Proteins of Immunological Interest, E. Kabat et al. 1983, U.S. Department of Health and Human Services). As noted above, the CDRs are primarily responsible for binding to an epitope of an antigen. An immune complex is an antibody, such as a monoclonal antibody, chimeric antibody, humanized antibody or human antibody, or functional antibody fragment, specifically bound to the antigen.

[0227] Chimeric antibodies are antibodies whose light and heavy chain genes have been constructed, typically by genetic engineering, from immunoglobulin variable and constant region genes belonging to different species. For example, the variable segments of the genes from a mouse monoclonal antibody can be joined to human constant segments, such as kappa and gamma 1 or gamma 3. In one example, a therapeutic chimeric antibody is a hybrid protein composed of the variable or antigen-binding domain from a mouse antibody and the constant or effector domain from a human antibody, although other mammalian species can be used, or the variable region can be produced by molecular techniques. Methods of making chimeric antibodies are well known in the art, e.g., see U.S. Patent No. 5,807,715. In one embodiment, the anti-DPP3 antibody according to the present invention is a chimeric antibody.

[0228] A “humanized” immunoglobulin is an immunoglobulin including a human framework region and one or more CDRs from a non-human (such as a mouse, rat, or synthetic) immunoglobulin. The non-human immunoglobulin providing the CDRs is termed a "donor" and the human immunoglobulin providing the framework is termed an "acceptor." In one embodiment, all the CDRs are from the donor immunoglobulin in a humanized immunoglobulin. Constant regions need not be present, but if they are, they must be substantially identical to human immunoglobulin constant regions, i.e., at least about 85- 90%, such as about 95% or more identical. Hence, all parts of a humanized immunoglobulin, except possibly the CDRs, are substantially identical to corresponding parts of natural human immunoglobulin sequences. A "humanized antibody" is an antibody comprising a humanized light chain and a humanized heavy chain immunoglobulin. A humanized antibody binds to the same antigen as the donor antibody that provides the CDR’s. The acceptor framework of a humanized immunoglobulin or antibody may have a limited number of substitutions by amino acids taken from the donor framework. Humanized or other monoclonal antibodies can have additional conservative amino acid substitutions, which have substantially no effect on antigen binding or other immunoglobulin functions. Exemplary conservative substitutions are those such as gly, ala; val, ile, leu; asp, glu; asn, gin; ser, thr; lys, arg; and phe, tyr.

[0229] Humanized immunoglobulins can be constructed by means of genetic engineering (e.g., see U.S. Patent No. 5,585,089). Human antibodies can also be prepared by using transgenic animals carrying a human immunoglobulin gene (for example, see WO 93 / 12227; WO 91 / 10741).

[0230] A human antibody is an antibody wherein the light and heavy chain genes are of human origin. Human antibodies can be generated using methods known in the art. Antibodies can be produced by immortalizing a (e.g. mammalian) B cell secreting the antibody of interest. Immortalization can be accomplished, for example, by Epstein-Barr Virus (EBV) infection of a B-cell or by fusing a B cell with a myeloma to make a hybridoma or fusing a B cell to a hybridoma cell to produce atrioma cell. Antibodies can also be produced by phage display methods (see, e.g., WO 91 / 17271; WO 92 / 001047; WO 92 / 20791) or selected from a combinatorial monoclonal antibody library (see for example the Morphosys website).

[0231] Thus, the anti-DPP3 antibody may have the formats known in the art. Examples are human antibodies, monoclonal antibodies, humanized antibodies, chimeric antibodies, CDR-grafted antibodies. In a preferred embodiment antibodies according to the present invention are recombinantly produced antibodies as e.g. IgG, a typical full-length immunoglobulin, or antibody fragments containing at least the variable domain of heavy and / or light chain as e.g. chemically coupled antibodies (fragment antigen binding) including but not limited to Fab-fragments including Fab minibodies, single chain Fab antibody, monovalent Fab antibody with epitope tags, e.g. Fab-V5Sx2; bivalent Fab (mini-antibody) dimerized with the CH3 domain; bivalent Fab or multivalent Fab, e.g. formed via multimerization with the aid of a heterologous domain, e.g. via dimerization of dHLX domains, e.g. Fab-dHLX-FSx2; F(ab‘)2-fragments, scFv-fragments, multimerized multivalent or / and multi-specific scFv-fragments, bivalent and / or bispecific diabodies, BITE® (bispecific T-cell engager), trifunctional antibodies, polyvalent antibodies, e.g. from a different class than G; single-domain antibodies, e.g. nanobodies derived from camelid or fish immunoglobulins and numerous others.

[0232] The characterization of the antibodies according to the present invention including epitope mapping, binding affinities, specificity, inhibitory potential is disclosed in more detail in Example 2.

[0233] In one embodiment said anti-DPP3 antibody or fragment thereof binding to DPP3 is an inhibitor of DPP3 activity.

[0234] The said anti-DPP3 antibody or fragment thereof binding to DPP3 inhibits the DPP3 activity of 50% or more, more preferred 60% or more, even more preferred 70% or more, most preferred 80% or more.

[0235] In a preferred embodiment the anti-DPP3 antibody format is selected from the group comprising Fv fragment, scFv fragment, Fab fragment, scFab fragment, F(ab)2 fragment and scFv-Fc Fusion protein. In another preferred embodiment the antibody format is selected from the group comprising scFab fragment, Fab fragment, scFv fragment and bioavailability optimized conjugates thereof, such as PEGylated fragments. One of the most preferred formats is the scFab format. In one embodiment of the invention anti-DPP3 antibodies according to the present invention may be produced as outlined in Example 1 by synthesizing fragments of DPP3 as antigens or full-length DPP3. Thereafter, antibodies to said fragments are identified using the below described methods or other methods as known in the art.

[0236] In particular embodiments, the fragments of DPP 3 antigen are a peptide, wherein the peptide is 15 to 55 amino acids long; is 15, 20, 25, 30, 35, 40, 45, 50, or 55 amino acids long; or is about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, or about 55 amino acids long, wherein the term “about” in this context of amino acid length for the peptide can be plus or minus 2 amino acids, e.g., about 20 amino acids means the range of 23-27 amino acids long.

[0237] In particular embodiments, the antigen comprises the peptide, and the peptide comprises the catalytic site from 316 to 669 amino acids, preferably including the catalytic motifs SEQ ID NO: 14 or SEQ ID NO: 15 and / or at least one or more of the important amino acids important for substrate binding and hydrolysis chosen from the list consisting of Glu316, Tyr318, Asp366, Asn391, Asn394, His568, Arg572, Arg577, Lys666, and Arg669.

[0238] Humanization of murine antibodies may be conducted according to the following procedure:

[0239] For humanization of an antibody of murine origin the antibody sequence is analyzed for the structural interaction of framework regions (FR) with the complementary determining regions (CDRs) and the antigen. Based on structural modelling an appropriate FR of human origin is selected and the murine CDR sequences are transplanted into the human FR. Variations in the amino acid sequence of the CDRs or FRs may be introduced to regain structural interactions, which were abolished by the species switch for the FR sequences. This recovery of structural interactions may be achieved by random approach using phage display libraries or via directed approach guided by molecular modelling (Almagro and Fransson 2008. Humanization of antibodies. Front Biosci. 13:1619-33}. (Almagro 2008) The use of molecular modeling to achieve a humanized antibodies may involve additional conservative amino acid substitutions at the junction of the CDR-FR. Exemplary conservative substitutions are those such as gly, ala; val, ile, leu; asp, glu; asn, gin; ser, thr; lys, arg; and phe, tyr. Additionally, back mutations may be implemented, where selection of residues outside of the region defining the specificity and thus target for back mutation (to mimic the source sequence, i.e., murine) to restore or improve the affinity of the humanized antibody. (Almagro 2008). When available, the three-dimensional models of the antibodies may be used to guide the humanization process.

[0240] The humanized anti-DPP3 antibody may recombinantly be produced with a CHO DG44 cell line. In a particular embodiment, the creation of a production cell, e.g., the CHO cell line encoding the DPP3 Binder, is used for producing the DPP3 Binder. The antibody binds to its target molecule DPP3, inhibiting its enzymatic activity as part of its mode of action. The humanized anti-DPP3 antibody is directed against a linear epitope SEQ ID NO: 3 (INPETG within the DPP3 sequence). The in silico derived molecular weights of the light and heavy chain of PCZ are 23,943 Dalton (Da) and 48,950 Da, respectively.

[0241] A unit dose in mg / kg refers to mg of antibody per kg of patient is based on a certain molecular weight (“MW”) of the administered antibody. All references to mg / kg in this application refer to the full-length anti-DPP3 antibody having a MW of 145,734 Da determined in silico. For any other MW of antibodies and fragments thereof the appropriate doses may be calculated accordingly

[0242] Corresponding molar doses of antibodies or antibody fragments having other molecular weights can in particular be calculated according to the following explanation:

[0243] A unit of dose in mg of antibody or fragment thereof per kg of patient is based on a specific molecular weight of the administered antibody or fragment thereof. If the molecular weight of the antibody would be for instance 145,734 g / mol (as is the case for the humanized anti-DPP3 antibody AK1967 (Procizumab; PCZ)), a dose of about 18 to 22 mg / kg would correspond to about 0.12 to 0.15 pmol / kg. If, in contrast, the molecular weight of the antibody would be for instance 165,000 g / mol, the same molar doses of about 0. 12 to 0. 15 pmol / kg would correspond to about 20 to 25 mg / kg.

[0244] The method of how the mass of PCZ was determined was as follows: The PCZ concentration attributed to the drug substance and drug product as well as the doses described in mass per weight (mg / kg) were based on measurement of the absorption of the PCZ at 280 nm measured in a spectrophotometer with a path length of 1 cm. Conversion of A280 nm measurement into mol / L was done by employing a theoretically calculated extinction coefficient sm„|;irusing the formula: molar Concentration=A280 nm / 8moiar. The extinction coefficient for the PCZ was calculated as follows: The amino acid sequences of the heavy and light chain of the PCZ, as shown below, were combined, duplicated, and then pasted in a calculator program (Expasy - ProtParam; https: / / web expasy.org / protparam / ; Protein Identification and Analysis Tools on the Expasy Server; Gasteiger E., Hoogland C., Gattiker A., Duvaud S., Wilkins M.R., Appel R.D., Bairoch A.; (In) John M. Walker (ed): The Proteomics Protocols Handbook, Humana Press (2005), pp. 571-607 Full text - Copyright Humana Press.):

[0245] PCZ heavy chain amino acid sequence (SEQ ID No. 34):

[0246] QITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEWLAHIWWNDNKSY NPALKSRLTITRDTSKNQVVLTMTNMDPVDTGTYYCARNYSYDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPP KPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPG

[0247] PCZ light chain amino acid sequence (SEQ ID No. 35)

[0248] DIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLYWYLQKPGQSPQLLIYKVSNRFSG VPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPWTFGGGTKVEIKRTVAAPSVFIFP PSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLS SP VTKSFNRGEC

[0249] The resulting calculation was:

[0250] Molar extinction coefficient sm„|;ir(at 280nm in H2O) = 228,440 M-1*cm-1

[0251] The value was calculated under the assumption that all cysteine residues form cystines. The molar concentration of PCZ can be determined by dividing the measured absorption at 280 by the molar extinction coefficient sm,:.|;ir.

[0252] For the conversion from mol into mass of PCZ a molecular weight of 145,734 g / mol was used. This molecular weight was calculated by amino acid sequence-based in silico prediction using the same calculator program as mentioned above (Expasy - ProtParam; https: / / web expasy.org / protparam / ): First, the amino acid sequence SEQ ID No. 34 of the heavy chain was predicted as having a molecular weight of 48,950 g / mol. Secondly, the amino acid sequence SEQ ID No. 35 of the light chain was predicted as having a molecular weight of 23,943 g / mol. The molecular weight for the full-length PCZ was then calculated as 2 x 48,950 Da + 2 x 23,943 Da = 145,734 g / mol (1 Da = 1 g / mol).

[0253] Of note, by using the method described above for the definition of the concentration of the PCZ, all concentrations and doses of the PCZ mentioned in the current invention are defined by the amino acid composition of the antibody only and do not take into consideration post-translational modifications such as glycosylation. Methods for obtaining monoclonal antibodies

[0254] In all of the following embodiments, the term “monoclonal antibody” is meant to include monoclonal antibodies, as well as fragments of monoclonal antibodies, such as the ones detailed herein, more particularly monoclonal antibodies.

[0255] Hybridoma

[0256] In a further aspect, the antibody according to the present invention is a monoclonal antibody obtainable by a method comprising: i) fusing antibody-secreting cells from an animal previously immunized with an antigen with myeloma cells to obtain a multitude of hybridomas, ii) isolating from said multitude of hybridomas a hybridoma producing a desired monoclonal antibody.

[0257] In certain embodiments, the antibody according to the present invention is a monoclonal antibody obtainable by isolating from a multitude of hybridomas a hybridoma producing a desired monoclonal antibody, wherein said multitude of hybridomas were produced by fusing antibody-secreting cells from an animal previously immunized with an antigen with myeloma cells to obtain multitude of hybridomas.

[0258] A desired monoclonal antibody is in particular a monoclonal antibody binding the antigen, in particular with a binding affinity of at least 107M1, preferred 108M1, more preferred affinity is greater than 109M1, most preferred greater than IO10M1.

[0259] In certain embodiments of the method for obtaining an antibody, in step i) the animal is a mammal, particularly a rabbit, a mouse or a rat, more particularly a mouse, more particularly a Balb / c mouse.

[0260] In certain embodiments of the method for obtaining an antibody, in step i) the antibody-secreting cell is a splenocyte, more particularly an activated B-cell.

[0261] In certain embodiments of the method for obtaining an antibody, in step i) fusing involves the use of polyethylene glycol.

[0262] In certain embodiments of the method for obtaining an antibody, in step i) the myeloma is derived from a mammal, in certain embodiments from the same species of mammal from which the multitude of antibody-secreting cells is obtained. In certain specific embodiments of the method for obtaining an antibody, in step i) the myeloma cells are of the cell line SP2 / 0. In certain embodiments of the method for obtaining an antibody, said fusing in step i) comprises PEG- assisted fusion, Sendai virus-assisted fusion or electric current-assisted fusion.

[0263] In certain embodiments of the method for obtaining an antibody, said isolating in step ii) comprises performing an antibody capture assay, an antigen capture assay, and / or a functional screen.

[0264] In certain embodiments of the method for obtaining an antibody, in step ii) isolating the hybridoma producing a desired monoclonal antibody may involve cloning and re-cloning the hybridomas using the limiting -dilution technique.

[0265] In one embodiment, said antibody capture assay comprises a) binding an antigen to a substrate, particularly a solid substrate, b) allowing the produced antibodies to bind to the antigen, c) removing unbound antibodies by washing, d) detecting bound antibodies.

[0266] In one embodiment, said antigen capture assay comprises a) binding the produced antibodies to a substrate, particularly a solid substrate, b) allowing antigen to bind to said antibodies, c) removing unbound antigen by washing, d) detecting bound antigen; or said antigen capture assay comprises a) allowing an antigen to bind the produced antibodies to form an antibody-antigen complex, b) binding said antibody-antigen complex to a substrate, particularly a solid substrate, c) removing unbound antigen by washing, d) detecting bound antigen.

[0267] It is apparent to the skilled person that in the context of the aforementioned antibody capture assay, the terms “substrate” and in particular “solid substrate” relate to a support, and in particular a solid support, i.e. a solid phase.

[0268] In one embodiment, said isolating of step ii) comprises performing an enzyme-linked immunosorbent assay, fluorescence-activated cell sorting, cell staining, immunoprecipitation, and / or a western blot.

[0269] In one embodiment, said detecting of the antibody or the antigen is accomplished with an immunoassay. In one embodiment, the animal is a transgenic animal, in particular a transgenic mouse (wherein in particular the mouse immunoglobulin (Ig) gene loci have been replaced with human loci within the transgenic animal genome), such as HuMabMouse or XenoMouse.

[0270] In one embodiment, the antigen comprises a peptide as described herein in Table 2, or Table 7 respectively, which in certain embodiments (in particular for immunization) may be conjugated to a protein, particularly a serum protein, more particularly a serum albumin, more particularly BSA.

[0271] In a preferred embodiment, the antibody according to the present invention is a monoclonal antibody obtainable by a method comprising: i) fusing splenocytes cells from a Balb / c mouse previously immunized with a peptide as described herein in Table 2 or Table 7 with SP2 / 0 myeloma cells using polyethylene glycol, to obtain a multitude of hybridomas, ii) isolating from said multitude of hybridomas a hybridoma producing a desired monoclonal antibody; more preferably, the method comprises

[0272] • growing hybridomas for a first period (in particular 2 weeks) in HAT medium [RPMI 1640 culture medium supplemented with 20% fetal calf serum and HAT-Supplement],

[0273] • followed replacing HAT medium with HT Medium for a multitude of passages (in particular 3),

[0274] • followed by returning to the normal cell culture medium for a second time period, in particular until the end of three weeks after fusion,

[0275] • primary screening of cell culture supernatants for antigen-specific IgG antibodies,

[0276] • propagating microcultures of cells that tested positive in 4),

[0277] • retesting cell culture supernatants of microcultures for antigen-specific IgG antibodies,

[0278] • cloning and re-cloning cultures that tested positive in 6), using the limiting-dilution technique,

[0279] • optionally determining the isotypes of clones obtained from 7),

[0280] • optionally purifying antibodies via Protein A.

[0281] Phage Display In a further aspect, the antibody according to the present invention is a monoclonal antibody obtainable by a method comprising: i) isolating at least one antibody having affinity to an antigen from an antibody gene library; ii) generating at least one cell strain expressing said at least one antibody; iii) isolating the at least one antibody from a culture of the at least one cell strain obtained in step ii).

[0282] An antibody having affinity to an antigen is in particular an antibody with a binding affinity of at least 107M1, preferred 108M1, more preferred affinity is greater than 109M1, most preferred greater than 101° M’1.

[0283] In a certain embodiment, the antibody according to the present invention is a monoclonal antibody obtainable by isolating at least one antibody from a culture derived from at least one cell strain which expressed at least one antibody having affinity to an antigen from an antibody gene library.

[0284] In one embodiment, the antigen comprises a peptide as described herein in Table 2, or Table 7 respectively, which in certain embodiments may be bound to a solid phase.

[0285] In certain embodiments of the method for obtaining an antibody, in step i) the antibody gene library is a naive antibody gene library, particularly a human naive antibody gene library, more particularly in said library the antibodies are presented via phage display, i.e. on phages comprising a nucleotide sequence encoding for such respective antibody; more particularly the library HAL 7, HAL 8, or HAL 9, more particularly a library comprising the human naive antibody gene libraries HAL7 / 8, or in alternative embodiments an equivalent of a library thereof listed herein.

[0286] In certain embodiments of the method for obtaining an antibody, in step i) screening comprises the use of an antigen, particularly an antigen containing a tag, more particularly a biotin tag, linked thereto via two different spacers. In particular embodiments, such panning strategy includes a mix of panning rounds with non-specifically bound antigen and antigen bound specifically via the tag, in the case of a biotin tag, bound to streptavidin. In this way, the background of non-specific binders may be minimized.

[0287] In certain embodiments of the method for obtaining an antibody, in step i), in embodiments wherein the library is a phage display library, the antibody is isolated by isolating a phage presenting said antibody (and comprising a nucleotide sequence encoding for the antibody).

[0288] In certain embodiments of the method for obtaining an antibody, in step ii) said cell strain is generated via introduction of a nucleotide sequence encoding for the antibody, in embodiments wherein the library in step i) is a phage display library, the isolated phage from step i) may be used to produce a bacterial strain, e.g. an Escherichia coli (E. coli) strain, expressing the antibody.

[0289] In certain embodiments of the method for obtaining an antibody, in step iv); in embodiments wherein the library in step i) is a phage display library and wherein a bacterial strain is produced in step ii), antibody may be isolated from the supernatant of the culture.

[0290] It is understood that, as used in describing the methods for obtaining an antibody, the term “one antibody” in the expression “at least one antibody” in particular may include more than one antibody molecule of antibodies having the same amino acid sequence. This understanding applies, mutatis mutandis, to the term “one cell strain”.

[0291] In certain embodiments of the method for obtaining an antibody, more than one antibody (referring to a multitude of antibodies having distinct amino acid sequences, respectively) is isolated in step i) and accordingly more than one cell strain is generated in step ii). Such method may involve the selection of clones that are positive for binding to the antigen, e.g. via a binding assay, e.g. an ELISA assay involving the antigen, and cells positive for binding to the antigen may be isolated to produce monoclonal cell strains.

[0292] In a preferred embodiment, the antibody according to the present invention is a monoclonal antibody obtainable by a method comprising: i) isolating at least one antibody having affinity to an antigen from an antibody gene library comprising the human naive antibody gene libraries HAL7 / 8, by eluting phages carrying said antibody from the library; ii) generating at least one bacterial strain, e.g., E. coli cell strain, expressing said at least one antibody; iii) isolating the at least one antibody from the supernatant a culture of the at least one E. coli cell strain obtained in step ii).

[0293] In a further aspect, an antibody fragment according to the present invention is produced by a method involving enzymatic digestion of an antibody. In certain embodiments, this method produces e.g. Fab or F(ab)2 antibody fragments. In certain embodiments, this method involves digestion with pepsin or papain, which are optionally immobilized on a surface.

[0294] In certain embodiments, antibodies may be humanized by CDR-grafting, in particular by a process involving the steps: extracting RNA from hybridomas expressing an antibody of interest (e.g. obtained by a method as described herein); amplifying said extracted RNA via RT-PCR, in particular with primer sets specific for the heavy and light chains of the antibody of interest, to obtain a DNA product; further amplifying said DNA product via PCR, in particular using semi-nested primer sets specific for antibody variable regions; determining the sequence of the DNA product; aligning said sequence with homologous human framework sequences to determine a humanized sequence for the variable heavy chain and the variable light chain sequences (of the desired antibody).

[0295] In certain embodiments, antibodies may be humanized by aligning the sequence of a DNA product that was obtained by amplifying RNA extracted from hybridomas expressing an antibody of interest via RT- PCR, in particular with primer sets specific for the heavy and light chains of the antibody of interest and further amplifying the DNA obtained therefrom via PCR, in particular using semi-nested primer sets specific for antibody variable regions, with homologous human framework sequences to determine a humanized sequence for the variable heavy chain and the variable light chain sequences (of the desired antibody).

[0296] In certain embodiments, antibodies may be humanized by determining the complementary determining regions (CDR), which may be accomplished by analysing the structural interaction of framework regions (FR) with the complementary determining regions (CDRs) and the antigen; transplanting said CDR sequences into a human framework region.

[0297] In certain embodiments, antibodies may be humanized by transplanting CDR sequences, which may preferably have been determined by analysing the structural interaction of framework regions (FR) with the complementary determining regions (CDRs) and the antigen, into a human framework region.

[0298] In certain embodiments variations in the amino acid sequence of the CDRs or FRs may be introduced to maintain structural interactions with the antigen (which may otherwise be abolished by introducing the human FR sequences), for instance by a random approach using phage display libraries or via directed approach guided by molecular modelling.

[0299] The DNA sequences encoding for antibodies determined as detailed herein can be transferred by known genetic engineering techniques into cells and used for production of the antibody. It is apparent to the skilled person that this can be accomplished transferring the DNA sequences encoding for antibodies determined as detailed herein by known recombinant genetic engineering techniques into expression vectors and introducing these into cells, and using such cells for production of the antibody, whereby the cell is a production cell.

[0300] Producing antibodies

[0301] In a further aspect, the antibody according to the present invention is a monoclonal antibody obtainable by the methods described herein, produced by a method comprising: culturing a cell strain comprising a nucleotide sequence encoding for the antibody; isolating the antibody from said culture.

[0302] In a further certain aspect, the antibody according to the present invention is a monoclonal antibody generated by the methods described herein, produced by isolating the antibody from a culture of a cell strain comprising a nucleotide sequence encoding for said antibody.

[0303] In certain embodiments of said method, the cell strain used to culture the antibody or antibody fragment thereof is produced as described herein above and may comprise bacterial cells, such as gram-negative bacteria, e.g. E. coli, Proteus mirabilis, or Pseudomonas putidas, gram-positive bacteria, e.g. Bacillus brevis, Bacillus subtilis, Bacillus megaterium, Lactobacilli such as Lactobacillus zeae / casei or Lactobacillus paracasei, or Streptomyces, such as Streptomyces lividans; eucariotic cells such as yest, e.g. Pichia pastoris, Saccharomyces cerevisiae, Hansenula polymorpha, Schizosaccharomyces pombe, Schwanniomyces occidentalis, Kluyveromyces lactis, or Yarrowia lipolytica; fugi, such as filamentous fungi, e.g. of the genus Trichoderma of Aspergillus, such as A. niger (e.g. subgenus A. awamori) and Aspergillus oryzae, Trichoderma reesei, Chrysosporium, such as C. lucknowense; protozoae, such as Leishmania, e.g. L. tarentolae; insect cells, such as insect cells transfected a Baculovirus, e.g. AcNPV, such as insect cell lines from Spodoptera frugiperda, e.g. Sf-9 or Sf-21, Drosophila melanogaster, e.g. DS2, or Trichopulsia ni, e.g. High Live cells (BTI-TN-5B1-4); mammalian cells such as hamster, e.g. Chinese hamster ovary such as K1-, DukX B11-, DG44, Lecl3, or BHK, mouse, e.g. mouse myeloma such as NSO, Homo sapiens, e.g. Per.C6, AGE1.HN, HEK293.

[0304] In certain embodiments of said method, the cells may be hybridoma cells, e.g. as described herein.

[0305] In a further aspect, procizumab (“PCZ”) is a monoclonal antibody obtainable by methods known in the art, or methods described herein. PCZ comprises two heavy chain sequences comprising SEQ ID NO: 34 and two light chain sequence comprising SEQ ID NO: 35. The respective SEQ ID NO: 12 for the heavy chain and SEQ ID NO: 13 for the light chain contain their respective N-terminal signal peptide sequences that can be used to manufacture the PCZ. The signal peptide sequence for the heavy chain is SEQ ID NO: 36 (MDPKGSLSWRILLFLSLAFELSYG) and for the light chain is SEQ ID NO: 37 (METDTLLLWVLLLWVPGSTG). As the PCZ is produced by the cell, the signal peptide sequence is cleaved off by cell before the protein is secreted. Culturing cells that express PCZ may be used to manufacture the protein at various scales. PCZ may be generated, for example, by any of the methods disclosed above used in the development and identification of the antibody. The DNA encoding the generated PCZ may be isolated and identified, for example, by DNA sequencing of the gene encoding the antibody against DPP3. Recombinant DNA technology well-known in the art may be used to manipulate the DNA. For example, the DNA encoding PCZ may be modified to make PCZ fragments and / or full-length PCZ to be combined with, for example, expression plasmid DNA. Further, the DNA can be codon optimized. The expression plasmid DNA encoding the PCZ or a fragment thereof may then be introduced, for example, by introducing the DNA, e.g., transfecting or in alternative embodiments transforming, into cells used for culturing and expressing the PCZ or a fragment thereof. The culturing cells expressing the PCZ or a fragment thereof may be used to manufacture the PCZ or a fragment thereof at various scales. The culturing cells may be, for example, Chinese Hamster Ovary (CHO) cells (including strains K1-, DukX B11-, DG44, Lecl3), Human Embryonic Kidney 293 (HEK293) cells, baby hamster kidney (BHK21) cells, murine myeloma cells (NSO and Sp2 / 0), Vero cells, MRC-5 cells, PerC.6 cells, AGE1.HN, or any other cells known in the art that are used to culture therapeutic proteins for manufacturing at various scales. Other non-mammalian cell lines or other cells may be used for culturing PCZ or a fragment thereof, such as, .S', cerevisiae and E. coli and the others described above. The post translational modification of the protein should be taken into account for the choice of the cells for culturing.

[0306] In certain embodiments of said method, culturing may take place in a static suspension culture, an agitated suspension culture, a membrane-based culture, a matrix-based culture or a high cell density bioreactor; a vessel for such culturing may be selected from the group comprising a T-flask, a roller culture, a spinner culture, a stirred tank bioreactor, an airlift bioreactor, a static membrane-based or matrix-based culture system, a suspension bioreactor, a fluidized bed bioreactor, a ceramic bioreactor, a perfusion system, a hollow fiber bioreactor.

[0307] In certain embodiments of said method, the cells may be immobilized on a matrix.

[0308] A high cell density bioreactor is in particular a culture system capable of generating cell densities greater than 108cells / ml.

[0309] In a further aspect, the antibody according to the present invention is a monoclonal antibody obtainable by the methods described herein, produced by a method comprising: generating a transgenic plant or animal comprising a nucleotide sequence encoding for the antibody; isolating the antibody from said plant or animal or a secretion or product of said plant or animal.

[0310] In a certain further aspect, the antibody according to the present invention is a monoclonal antibody obtainable by the methods described herein, produced by isolating the antibody from a transgenic plant or transgenic animal or a secretion or product of a transgenic plant or transgenic animal having a nucleotide sequence encoding for the antibody.

[0311] Said animal may e.g. be selected from a chicken, a mouse, a rat, a rabbit, a cow, a goat, a sheep, a pig; said secretion or product may e.g. be milk or an egg. Said plant may e.g. be selected from tobacco (N. tabacum or N. benthamiana), duckweed (Lemna minor), Chlamydomonas reinhardtii, rice, Arabidopsis thaliana, alfalfa (Medicago sativa), lettuce, maize.

[0312] The antibodies can in certain embodiments be isolated by physicochemical fractionation, e.g. size exclusion chromatography, precipitation, e.g. using ammonium sulphate, ion exchange chromatography, immobilized metal chelate chromatography gel fdtration, zone electrophoresis; based on their classification e.g. binding to bacterial proteins A, G, or L, jacalin; antigen-specific affinity purification via immobilized ligands / antigens; if necessary, low molecular weight components can be removed by methods like dialysis, desalting, and diafiltration.

[0313] In some embodiments the antibody is encoded by a nucleotide sequence where the nucleotide sequence is a reverse transcription of an amino acid sequence from an antibody produced by one of the processes described herein.

[0314] A person of ordinary skill in the art would appreciate that the biosimilarity of an antibody to an anti- DPP3 antibody or fragment thereof claimed herein can be determined by, for example, following the U.S. FDA’s Guidance “Questions and Answers on Biosimilar Development and the BPCI Act” dated September 2021, incorporated in full herein.

[0315] In another preferred embodiment, the anti-DPP3 antibody, anti-DPP3 antibody fragment, or anti-DPP3 non-Ig scaffold is a full-length antibody, antibody fragment, or non-Ig scaffold.

[0316] In certain embodiments of the invention, the DPP3 binder is an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold that binds an epitope of at least 4 or 5 amino acids in length comprised in SEQ ID NO: 1. In one embodiment said binder is an anti-DPP3 antibody or anti- DPP3 antibody fragment or anti-DPP3 non-Ig scaffold that binds an epitope of at least 4 amino acids in length comprised in SEQ ID NO: 1. In another embodiment said binder is an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold that binds an epitope of at least 5 amino acids in length comprised in SEQ ID NO: 1.

[0317] In certain embodiments of the invention, the DPP3 binder is an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold that binds an epitope of at least 4 or 5 amino acids in length comprised in SEQ ID NO: 2, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO: 1.

[0318] In certain embodiments of the invention, the DPP3 binder is an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold that binds an epitope of at least 4 amino acids in length comprised in SEQ ID NO: 2, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO:

[0319] 1. In another embodiment said binder is an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti- DPP3 non-Ig scaffold that binds an epitope of at least 5 amino acids in length comprised in SEQ ID NO:

[0320] 2, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO: 1.

[0321] In certain embodiments of the invention, the DPP3 binder is an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold that binds an epitope of at least 4 or 5 amino acids in length comprised in SEQ ID NO: 3, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO: 1. In one embodiment said binder is an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold that binds an epitope of at least 4 amino acids in length comprised in SEQ ID NO: 3, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO: 1. In another embodiment said binder is an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold that binds an epitope of at least 5 amino acids in length comprised in SEQ ID NO: 3, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO: 1.

[0322] An epitope, also known as antigenic determinant, is the part of an antigen that is recognized by the immune system, specifically by antibodies. For example, the epitope is the specific piece of the antigen to which an antibody binds. The part of an antibody that binds to the epitope is called a paratope. The epitopes of protein antigens are divided into two categories, conformational epitopes and linear epitopes, based on their structure and interaction with the paratope. Conformational and linear epitopes interact with the paratope based on the 3-D conformation adopted by the epitope, which is determined by the surface features of the involved epitope residues and the shape or tertiary structure of other segments of the antigen.

[0323] A conformational epitope is formed by the 3-D conformation adopted by the interaction of discontinuous amino acid residues. A linear or a sequential epitope is an epitope that is recognized by antibodies by its linear sequence of amino acids, or primary structure and is formed by the 3-D conformation adopted by the interaction of contiguous amino acid residues.

[0324] In specific embodiments of the invention the antibody or fragment thereof is a monoclonal antibody or a fragment thereof.

[0325] In one embodiment of the invention the anti-DPP3 antibody or the anti-DPP3 antibody fragment is a human or humanized antibody or derived therefrom. In one specific embodiment one or more (murine) CDR’s are grafted into a human antibody or antibody fragment.

[0326] In one embodiment of the invention the antibody is a human or humanized CDR-grafted antibody or antibody fragment thereof that binds to DPP3, wherein the human or humanized CDR-grafted antibody or antibody fragment thereof comprises an antibody heavy chain (H chain) comprising SEQ ID No. 7: GFSLSTSGMS

[0327] SEQ ID No. 8: IWWNDNK

[0328] SEQ ID No. 9: ARNYSYDY and / or further comprises an antibody light chain (L chain) comprising:

[0329] SEQ ID No. 10: RSLVHSIGSTY

[0330] CDR2: KVS

[0331] SEQ ID No. 11: SQSTHVPWT or in particular embodiments a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0332] In one embodiment of the invention, the antibody is a human or humanized monoclonal antibody that binds to DPP3 or an antibody fragment thereof that binds to DPP3 wherein the heavy chain comprises at least one CDR selected from the group comprising:

[0333] SEQ ID No. 7: GFSLSTSGMS

[0334] SEQ ID No. 8: IWWNDNK

[0335] SEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR selected from the group comprising:

[0336] SEQ ID No. 10: RSLVHSIGSTY

[0337] CDR2: KVS

[0338] SEQ ID No. 11: SQSTHVPWT or in particular embodiments a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%. In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein the heavy chain comprises the CD Rs selected from the group comprising:

[0339] SEQ ID No. 7: GFSLSTSGMS,

[0340] SEQ ID No. 8: IWWNDNK and / or

[0341] SEQ ID No. 9: ARNYSYDY or in particular embodiments a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0342] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein the light chain comprises the CD Rs selected from the group comprising:

[0343] SEQ ID No. 10: RSLVHSIGSTY,

[0344] CDR2: KVS and / or

[0345] SEQ ID No. 11: SQSTHVPWT or in particular embodiments a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%..

[0346] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein the heavy chain comprises the CD Rs selected from the group comprising:

[0347] SEQ ID No. 7: GFSLSTSGMS,

[0348] SEQ ID No. 8: IWWNDNK and / or

[0349] SEQ ID No. 9: ARNYSYDY and wherein the light chain comprises the CDRs selected from the group comprising:

[0350] SEQ ID No. 10: RSLVHSIGSTY,

[0351] CDR2: KVS and / or

[0352] SEQ ID No. 11: SQSTHVPWT or in particular embodiments a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0353] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a heavy chain Fab fragment comprising SEQ ID NO: 16.

[0354] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a heavy chain Fab fragment comprising SEQ ID NO: 38.

[0355] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a light chain Fab fragment comprising SEQ ID NO: 13.

[0356] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a light chain Fab fragment comprising SEQ ID NO: 35.

[0357] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a heavy chain Fab fragment comprising SEQ ID NO: 16 and comprises a light chain Fab fragment comprising SEQ ID NO: 13.

[0358] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a heavy chain Fab fragment comprising SEQ ID NO: 38 and comprises a light chain Fab fragment comprising SEQ ID NO: 35.

[0359] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a heavy chain variable region portion from SEQ ID NO: 16.

[0360] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a heavy chain variable region portion from SEQ ID NO: 38. In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a light chain variable region portion from SEQ ID NO: 13.

[0361] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a light chain variable region portion from SEQ ID NO: 35.

[0362] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a heavy chain variable region portion from SEQ ID NO: 16 and comprises a light chain variable region portion from SEQ ID NO: 13.

[0363] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a heavy chain variable region portion from SEQ ID NO: 38 and comprises a light chain variable region portion from SEQ ID NO: 35.

[0364] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a single chain F-Variable domain (ScFv) and wherein the ScFv comprises a heavy chain variable region portion from SEQ ID NO: 16 and comprises a light chain variable region portion from SEQ ID NO: 13, and wherein the heavy chain variable region portion from SEQ ID NO: 16 and a light chain variable region portion from SEQ ID NO: 13 are linked together via linker.

[0365] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a ScFv and wherein the ScFv comprises a heavy chain variable region portion from SEQ ID NO: 38 and comprises a light chain variable region portion from SEQ ID NO: 35, and wherein the heavy chain variable region portion from SEQ ID NO: 38 and a light chain variable region portion from SEQ ID NO: 35 are linked together via linker.

[0366] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a Fab fragment and wherein the Fab fragment comprises a heavy chain Fab fragment comprising SEQ ID NO: 16 and comprises a light chain Fab fragment comprising SEQ ID NO: 13.

[0367] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a Fab fragment and wherein the Fab fragment comprises a heavy chain Fab fragment comprising SEQ ID NO: 38 and comprises a light chain Fab fragment comprising SEQ ID NO: 35.

[0368] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a heavy chain variable region comprising SEQ ID NO: 5.

[0369] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a light chain variable region comprising SEQ ID NO: 6.

[0370] In one embodiment of the present invention, the antibody is an anti -DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof comprises a heavy chain variable region comprising SEQ ID NO: 5 and comprises a light chain variable region comprising SEQ ID NO: 6.

[0371] The anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold according to the present invention exhibits an affinity towards human DPP3 in such that affinity constant is greater than 10'7M, preferred IO-8M, preferred affinity is greater than 10'9M, most preferred higher than IO-10M. A person skilled in the art knows that it may be considered to compensate lower affinity by applying a higher dose of compounds and this measure would not lead out-of-the-scope of the invention.

[0372] A further embodiment of the present invention is a monoclonal antibody or fragment that binds to DPP3 for the treatment of shock in a patient having shock, wherein said patient is an extreme-critically ill patient, wherein said antibody or fragment comprises the following sequence:

[0373] SEQ ID No. 12

[0374] MDPKGSLSWRILLFLSLAFELSYGQITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQ PPGKALEWLAHIWWNDNKSYNPALKSRLTITRDTSKNQVVLTMTNMDPVDTGTYYCARNY SYDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQS SGLYSLS S VVTVPS S SLGTQTYICNVNHKP SNTKVDKKVEPKSCDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG and / or

[0375] SEQ ID No. 13

[0376] METDTLLLWVLLLWVPGSTGDIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLYWYLQ KPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPWTFGGG TKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC or in particular embodiments sequences that are > 80% identical, preferably > 90% identical, preferably > 95% identical, preferably > 98% identical, preferably > 99% identical thereto, wherein the heavy chain comprises at least one CDR selected from the group comprising SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or

[0377] SEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR selected from the group comprising: SEQ ID No. 10: RSLVHSIGSTY, CDR2: KVS and / or

[0378] SEQ ID No. 11: SQSTHVPWT, or a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences, provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0379] In one embodiment of the invention said anti-DPP3 antibody comprises the following sequences: SEQ ID No. 12

[0380] MDPKGSLSWRILLFLSLAFELSYGQITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQ PPGKALEWLAHIWWNDNKSYNPALKSRLTITRDTSKNQVVLTMTNMDPVDTGTYYCARNY SYDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQS SGLYSLS S VVTVPS S SLGTQTYICNVNHKP SNTKVDKKVEPKSCDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG and / or

[0381] SEQ ID No. 13 METDTLLLWVLLLWVPGSTGDIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLYWYLQ KPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPWTFGGG TKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC or in particular embodiments sequences that are > 80% identical, preferably > 90% identical, preferably > 95% identical, preferably > 98% identical, preferably > 99% identical thereto, wherein the heavy chain comprises at least one CDR selected from the group comprising SEQ ID No. 7: GFSLSTSGMS,

[0382] SEQ ID No. 8: IWWNDNK and / or

[0383] SEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR selected from the group comprising: SEQ ID No. 10: RSLVHSIGSTY,

[0384] CDR2: KVS and / or

[0385] SEQ ID No. 11: SQSTHVPWT, or a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences, provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0386] In one embodiment of the present invention, the antibody is an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said antibody or fragment thereof is PCZ.

[0387] A further embodiment of the present invention is a monoclonal antibody or fragment that binds to DPP3 for the treatment of shock in a patient having shock, wherein said patient is an extreme-critically ill patient, wherein said antibody or fragment comprises the following sequence: SEQ ID No. 34

[0388] QITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEWLAHIWWNDNKSYNP ALKSRLTITRDTSKNQVVLTMTNMDPVDTGTYYCARNYSYDYWGQGTLVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV PSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW

[0389] LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPG and / or SEQ ID No. 35

[0390] DIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLYWYLQKPGQSPQLLIYKVSNRFSGVPD RFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLS SP VTKSFNRGEC or in particular embodiments sequences that are > 80% identical, preferably > 90% identical, preferably > 95% identical, preferably > 98% identical, preferably > 99% identical thereto, wherein the heavy chain comprises at least one CDR selected from the group comprising SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or

[0391] SEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR selected from the group comprising: SEQ ID No. 10: RSLVHSIGSTY,

[0392] CDR2: KVS and / or

[0393] SEQ ID No. 11: SQSTHVPWT, or a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences, provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0394] In one embodiment of the invention said anti-DPP3 antibody comprises the following sequences:

[0395] SEQ ID No. 34

[0396] QITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEWLAHIWWNDNKSYNP ALKSRLTITRDTSKNQVVLTMTNMDPVDTGTYYCARNYSYDYWGQGTLVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV PSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW

[0397] LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPG and / or

[0398] SEQ ID No. 35

[0399] DIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLYWYLQKPGQSPQLLIYKVSNRFSGVPD RFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLS SP VTKSFNRGEC or in particular embodiments sequences that are > 80% identical, preferably > 90% identical, preferably > 95% identical, preferably > 98% identical, preferably > 99% identical thereto, wherein the heavy chain comprises at least one CDR selected from the group comprising SEQ ID No. 7: GFSLSTSGMS,

[0400] SEQ ID No. 8: IWWNDNK and / or

[0401] SEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR selected from the group comprising: SEQ ID No. 10: RSLVHSIGSTY,

[0402] CDR2: KVS and / or

[0403] SEQ ID No. 11: SQSTHVPWT, or a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences, provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0404] This means, in one embodiment of the invention, the CDRs do not exhibit any variations of the sequence. Any variation of the above sequence is outside of the CDRs in said embodiment.

[0405] Dosage and Formulation

[0406] Dosage regimens are determined and adjusted to provide the optimum desired response (e.g., a therapeutic response).

[0407] In one embodiment of the invention said anti-DPP3 antibody is administered at a dose between 1 to 30 mg / kg bodyweight, preferably 2 to 20 mg / kg bodyweight, preferably 3 to 15 mg / kg body weight, preferably 5 to 12 mg / kg bodyweight, more preferably 7 to 10 mg / kg bodyweight, and most preferably 10 mg / kg bodyweight.

[0408] In a further embodiment of the invention said anti-DPP3 antibody is administered intravenously at a dose of 10 mg / kg bodyweight, after which it is titrated up to no greater than 30 mg / kg bodyweight or titrated down to no less than 1 mg / kg bodyweight. In one embodiment of the invention said anti-DPP3 antibody is administered at a dose of 5 to 12 mg / kg bodyweight, wherein said anti-DPP3 antibody is subsequently administered in a plurality of additional doses and wherein said additional doses are no greater than 30 mg / kg bodyweight and no less than 1 mg / kg bodyweight. In another embodiment the anti-DPP3 antibody is administered intravenously at a dose no greater than 30 mg / kg bodyweight to no less than 1 mg / kg bodyweight to a patient suffering from shock wherein said patient is undergoing treatment with one or more vasopressors. In one embodiment of the invention said anti-DPP3 antibody is applied as infusion over 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours.

[0409] In one embodiment of the invention an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody is administered at a dose of 10 mg / kg bodyweight and applied as infusion over 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours. In another embodiment the anti-DPP3 antibody is administered intravenously at a dose no greater than 30 mg / kg bodyweight to no less than 1 mg / kg bodyweight for a period of up to 3 hours. In one embodiment of the invention said anti-DPP3 antibody is administered intravenously at a dose of 10 mg / kg bodyweight, after which said dose is titrated up to no greater than 30 mg / kg bodyweight or titrated down to no less than 1 mg / kg bodyweight to achieve a MAP of 65 mmHg in a patient suffering from shock.

[0410] It is apparent that particular embodiments of the invention involve an anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to the invention, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody is administered at a dose of 10 mg / kg bodyweight and administered as infusion over 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours.

[0411] The term “pharmaceutical formulation” or “dosage form” means a pharmaceutical ingredient in combination with at least one pharmaceutically acceptable excipient, which is in such form as to permit the biological activity of a pharmaceutical ingredient contained therein to be effective, and which contains no additional components which are unacceptably toxic to a subject to which the formulation would be administered. The term "pharmaceutical ingredient" means a therapeutic component (e.g., an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold) which can be combined with pharmaceutically acceptable excipients to provide a pharmaceutical formulation or dosage form.

[0412] In one embodiment the pharmaceutical formulation suitable for infusion or injection is an aqueous solution comprising a buffer. In another embodiment such solution comprises a sugar, wherein the sugar is sucrose, trehalose, or mannitol. In a further embodiment such solution comprises an amino acid, wherein the amino acid is lysine, arginine, or histidine. In one embodiment the pharmaceutical formulation suitable for injection is a dispersion comprising glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof. Pharmaceutical formulation solutions are sterilized prior to administration to a patient. In one embodiment, the anti-DPP3 antibody is contained in a pharmaceutical formulation. In one embodiment, the anti-DPP3 antibody pharmaceutical formulation is a concentrated solution (i.e., to be diluted prior to administration) for infusion and contains the anti-DPP3 antibody as an active substance together with one or more pharmaceutically acceptable excipients. The pharmaceutically acceptable excipients are those that are compatible with the anti-DPP3 antibody. The pharmaceutically acceptable excipients for use with the anti-DPP3 antibody include stabilizers, antioxidants, tonicity modifiers, and buffers. Suitable stabilizers include sugars, sugar alcohols, amino acids, surfactants, polyols, and stabilizing proteins. Examples of sugar stabilizers are sucrose, glucose, lactose, trehalose, and maltose; examples of sugar alcohol stabilizers include mannitol, xylitol, and sorbitol; examples of amino acid stabilizers include arginine, histidine, glycine, proline, alanine, lysine, leucine, and methionine; and examples of surfactant stabilizers include Polysorbate 20 and Polysorbate 80 (i.e., Tween 20 and Tween 80, respectively), Poloxamers (i.e., poloxamer 188 and Pluronic F68, 127), and PEGylated surfactants; examples of stabilizing polyols include glycerol, polyethylene glycol (PEG, low MW) and examples of stabilizing proteins include transferrin, human serum albumin, and bovine serum albumin. Antioxidants useful in an anti-DPP3 concentrated solution include methionine, ascorbic acid, reduced glutathione, cysteine, Thio glycerol, sodium thiosulfate, platinum, N-acetyl-DL-tryptophan, ethylenediaminetetraacetic acid (ED TA) and diethylenetriaminepentaacetic acid (DTP A). Examples of suitable tonicity modifiers include sodium chloride, potassium chloride, calcium chloride, mannitol, sorbitol, and glycerol. Buffers suitable in a concentrated solution include histidine, citrate, phosphate (e.g., phosphate-buffered saline), acetate, succinate, MES, HEPES, and Tris (e.g. tris-buffered saline). Examples of suitable pHs are in the range of 4.8 to 8.0, preferred in the range of 5.0 to 6.5, or more preferred in the range of 5.5 to 6.8; or at a pH of about 5.0, about 5.5, about 6.0, or about 6.5; preferred at a pH of 6.5. In one embodiment the anti-DPP3 antibody is lyophilized and is formulated with bulking agents (i.e., lyoprotectants) and suitable bulking agents include sucrose, trehalose, mannitol, glycine, and dextran. The excipients can serve multiple purposes within a single formulation and are not limited to a single function. Multiple excipients can be included in the formulation to fulfill similar purposes. In one embodiment, the anti-DPP3 antibody is formulated with 10 mM Histidine-HCl, 250 mM Trehalose, and 20 mM L-Methionine pH 6.5 buffer.

[0413] In one embodiment, the anti-DPP3 antibody pharmaceutical formulation may be stored at 2-8 °C. Before use, the vials of the concentrated solution are equilibrated to room temperature and then diluted with 0.9% saline, water for injection (WFI), phosphate buffered with saline (PBS), or WFI formulated with a tonicity modifier(s) (e.g., sodium chloride, potassium chloride, calcium chloride, mannitol, sorbitol, and glycerol) prior to infusion. In one embodiment the diluent has the same or equivalent excipient composition as is used for the concentrated pharmaceutical form, albeit, without the anti-DPP3 antibody. In another embodiment the diluent has the same or equivalent excipient composition and concentrations of the excipients as used for the concentrated pharmaceutical form, albeit, without the anti-DPP3 antibody.

[0414] In one embodiment the pharmaceutical formulation is in a freeze-dried state. Such a freeze-dried state may be a lyophilized cake that can be reconstituted to create a concentrated solution or a solution suitable for infusion or injection as described above.

[0415] The anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold can be administered alone or with other pharmaceutical ingredients. For example, the anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold can be administered before, substantially contemporaneous with, and / or after another pharmaceutical ingredient. The one or more pharmaceutical ingredients can be combined in a single pharmaceutical formulation. Other pharmaceutical ingredients include those considered to be the standard of care treatment for shock including but not limited to vasopressors and inotropic agents. Examples of vasopressors include vasopressin, norepinephrine, and epinephrine. Examples of inotropic agents include dopamine and dobutamine.

[0416] The pharmaceutical formulation comprising the anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold may be used to alleviate various conditions associated with shock, including but not limited to inadequate perfusion, inadequate tissue oxygenation, and impaired organ function. Such pharmaceutical formulation may be used to decrease blood cDPP3 activity levels, to decrease blood lactate levels, decrease blood IL-6 levels, decrease blood NT-proBNP levels, decrease blood creatinine levels, and increase glomerular filtration rate in a patient suffering from shock. Administration of the pharmaceutical formulation comprising the anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold to a patient suffering from shock may be used to achieve or maintain a MAP of 65 mmHg. Moreover, administration of the pharmaceutical formulation comprising the anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold to a patient in refractory shock wherein said patient is undergoing treatment with one or more vasopressors may be used to reduce the dose of said vasopressors.

[0417] One embodiment of the present invention is a pharmaceutical formulation for use in the treatment of shock in a patient having shock, wherein said patient is an extreme-critically ill patient and wherein said pharmaceutical composition comprises an antibody or fragment or scaffold according to the present invention.

[0418] One embodiment of the present invention is a pharmaceutical formulation for use in the treatment of shock in a patient having shock, wherein said patient is an extreme-critically ill patient according to the present invention and wherein said pharmaceutical formulation is a solution, preferably a ready-to-use solution.

[0419] One embodiment of the present invention is a pharmaceutical formulation for use in the treatment of shock in a patient having shock, wherein said patient is an extreme-critically ill patient according to the present invention and wherein said pharmaceutical formulation is in a freeze-dried state. Such a freeze- dried state may be a lyophilized cake that can be reconstituted using sterile water or a buffer.

[0420] In another embodiment of the present invention said pharmaceutical composition that is to be used in the treatment of shock in a patient having shock, wherein said patient is an extreme-critically ill patient according to the present invention is administered orally, percutaneously, subcutaneously, intradermally, sublingually, intramuscularly, intraarterially, intracerebrally, intracerebroventricularly, intravenously, or via the central nervous system (CNS) or via intraperitoneal administration. More specifically, said pharmaceutical composition may be administered by continuous intravenous infusion, a single subcutaneous bolus, or several divided doses administered intravenously or subcutaneously over time.

[0421] Another embodiment of the present invention is a kit comprising the herein disclosed anti-DPP3 antibody or an anti-DPP3 antibody fragment for use in the treatment of shock in a patient having shock, wherein said patient is an extreme-critically ill patient according to the present invention. With the above context, the following consecutively numbered embodiments provide further specific aspects of the invention:

[0422] 1. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said patient is an extreme-critically ill patient.

[0423] 2. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 1, wherein said shock is selected from the group comprising hypovolemic shock, cardiogenic shock, obstructive shock and distributive shock.

[0424] 3. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiments 1 or 2 wherein said shock patient has combined cardiogenic and septic shock.

[0425] 4. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of a patient having shock in a patient having shock according to embodiments 1 to 3, wherein said shock is refractory shock. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiments 1 to 4, wherein said extreme-critically ill patient has a 24- hour probability of mortality of at least 70%, preferably of at least 80%, more preferably of at least 90%, most preferably of at least 95% and / or has organ dysfunction requiring mechanical or pharmacological support of at least two organs. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 5, wherein said organ dysfunction is defined as cardiovascular dysfunction, respiratory dysfunction (lung), kidney dysfunction, liver dysfunction, central nervous system dysfunction or dysfunction of the coagulation system. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiments 5 or 6, wherein said organ support is selected from kidney support, respiratory (lung) support, cardiovascular support and / or liver support. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiments 1 to 7, wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold and wherein said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 8, wherein said threshold ofDPP3 concentration is between 20 and 120 ng / mL, more preferred between 25 and 100 ng / mL, even more preferred between 30 and 80 ng / mL, even more preferred between 35 and 60 ng / mL, even more preferred between 40 and 50 ng / ml most preferred said threshold is 40 ng / mL. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 8, wherein said threshold of enzymatic activity of DPP3 is between 1.76 and 9.68 pmol / (min*L), more preferred between 2.15 and 8.09 pmol / (min*L), even more preferred between 2.55 and 6.51 pmol / (min*L), even more preferred between 2.94 and 4.93 pmol / (min*L), even more preferred between 3.34 and 4.13 pmol / (min*L), most preferred said threshold is 3.34 pmol / (min*L). Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiment 7 to 10, wherein said bodily fluid is selected from the group comprising whole blood, serum and plasma. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiments 1 to 11, wherein the patient is characterized by a level of lactate of 2 mmol / 1 or higher and / or a level of IL-6 of 5.9 pg / ml or higher and / or a level of CRP of 1 mg / dL or higher and / or a level of PCT of 0.5 ng / ml or higher and / or a level of bio- ADM of 70 pg / ml or higher and / or serum creatinine of 1.18 mg / dL (for men) and 1.02 mg / dL (for women) and / or a supranormal level of NT-proBNP in a sample of bodily fluid of said patient. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiments 1 to 12, wherein an anti-DPP3 antibody is used. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiments 1 to 13, wherein an anti-DPP3antibody is used and wherein the heavy chain comprises at least one CDR, preferably at least three CDRs selected from the group comprising:

[0426] SEQ ID No. 7: GFSLSTSGMS,

[0427] SEQ ID No. 8: IWWNDNK and / or

[0428] SEQ ID No. 9: ARNYSYDY. and wherein the light chain comprises at least one CDR, preferably at least three CDRs selected from the group comprising:

[0429] SEQ ID No. 10: RSLVHSIGSTY,

[0430] CDR2: KVS and / or

[0431] SEQ ID No. 11: SQSTHVPWT or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%. Anti-DPP3 antibody thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiments 1 to 14, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody comprises the following sequences:

[0432] SEQ ID No. 12

[0433] MDPKGSLSWRILLFLSLAFELSYGQITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSV GWIRQPPGKALEWLAHIWWNDNKSYNPALKSRLTITRDTSKNQVVLTMTNMDPVDT GTYYCARNYSYDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT KVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK VSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPG and / or

[0434] SEQ ID No. 13

[0435] METDTLLLWVLLLWVPGSTGDIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLY WYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQST HVPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC or alternatively sequences that are > 80% identical, preferably > 90% identical, preferably > 95% identical, preferably > 98% identical, preferably > 99% identical thereto, wherein the heavy chain comprises at least one CDR selected from the group comprising SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or

[0436] SEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR selected from the group comprising: SEQ ID No. 10: RSLVHSIGSTY, CDR2: KVS and / or

[0437] SEQ ID No. 11: SQSTHVPWT, or a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences, provided that the binding affinity to DPP3 is diminished by not more than 20%. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiments 1 to 15, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3antibody is administered at a dose between 1 to 30 mg / kg bodyweight, preferably 2 to 20 mg / kg bodyweight, preferably 3 to 15 mg / kg bodyweight, preferably 5 to 12 mg / kg body weight, preferably 7 to 10 mg / kg bodyweight and most preferred 10 mg / kg bodyweight. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiments 1 to 16, wherein an anti-DPP3antibody is used and wherein said anti-DPP3 is administered for 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiments 1 to 17, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody is administered at a dose of 10 mg / kg bodyweight and administered as infusion over 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 1 to 18, wherein the heavy chain comprises the CDRs selected from the group comprising:

[0438] SEQ ID No. 7: GFSLSTSGMS,

[0439] SEQ ID No. 8: IWWNDNK and / or

[0440] SEQ ID No. 9: ARNYSYDY or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, according to embodiment 1 to 19, wherein the light chain comprises the CDRs selected from the group comprising:

[0441] SEQ ID No. 10: RSLVHSIGSTY,

[0442] CDR2: KVS and / or

[0443] SEQ ID No. 11: SQSTHVPWT or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 1 to 20, wherein the heavy chain comprises the CDRs selected from the group comprising:

[0444] SEQ ID No. 7: GFSLSTSGMS,

[0445] SEQ ID No. 8: IWWNDNK and / or SEQ ID No. 9: ARNYSYDY. and wherein the light chain comprises the CDRs selected from the group comprising:

[0446] SEQ ID No. 10: RSLVHSIGSTY,

[0447] CDR2: KVS and / or

[0448] SEQ ID No. 11: SQSTHVPWT or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiments 1 to 21, wherein said antibody or fragment thereof comprises a heavy chain Fab Fragment comprising SEQ ID NO: 16. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiments 1 to 22, wherein said antibody or fragment thereof comprises a light chain Fab Fragment comprising SEQ ID NO: 13. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 1 to 23, wherein said antibody or fragment thereof comprises a heavy chain Fab Fragment comprising SEQ ID NO: 16 and comprises a light chain Fab Fragment comprising SEQ ID NO: 13. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 1 to 24, wherein said antibody or fragment thereof comprises a heavy chain variable region portion of SEQ ID NO: 16. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 1 to 25, wherein said antibody or fragment thereof comprises a light chain variable region portion of SEQ ID NO: 13. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 1 to 26, wherein said antibody or fragment thereof comprises a heavy chain variable region portion of SEQ ID NO: 16 and comprises a light chain variable region portion of SEQ ID NO: 13. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 1 to 27, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a ScFv and wherein the ScFv comprises a heavy chain variable region portion of SEQ ID NO: 16 and comprises a light chain variable region portion of SEQ ID NO: 13 portion of SEQ ID NO: 13 and wherein the ScFv further comprises a linker connecting the heavy chain variable region portion of SEQ ID NO: 16 and the light chain variable region portion of SEQ ID NO: 13. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 1 to 28, wherein said antibody or fragment thereof comprises a heavy chain Fab region comprising SEQ ID NO: 16 and comprises a light chain variable region comprising SEQ ID NO: 13. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 1 to 29, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a ScFv and wherein the ScFv comprises a heavy chain variable region comprising SEQ ID NO: 16 and comprises a light chain variable region comprising SEQ ID NO: 13. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 1 to 30, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a Fab fragment and wherein the Fab fragment comprises a heavy chain Fab region comprising SEQ ID NO: 16 and comprises a light chain variable region comprising SEQ ID NO: 13. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 1 to 31, wherein said antibody or fragment thereof comprises a heavy chain variable region comprising SEQ ID NO: 5. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiment 1 to 32, wherein said antibody or fragment thereof comprises a light chain variable region comprising SEQ ID NO: 6. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 1 to 33, wherein said antibody or fragment thereof comprises a heavy chain variable region comprising SEQ ID NO: 5 and comprises a light chain variable region comprising SEQ ID NO: 6. A method for the treatment of shock in an extreme- critically ill patient having shock, comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof according to any of embodiments 1 to 34; wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold of DPP3 concentration; wherein said threshold is between 20 and 120 ng / mL; and wherein said anti-DPP3 antibody is administered at a dose between 1 and 30 mg / kg bodyweight. The method of embodiment 35, wherein said threshold is between 25 and 100 ng / mL. The method of embodiment 36, wherein said threshold is between 30 and 80 ng / mL. The method of embodiment 37, wherein said threshold is between 35 and 60 ng / mL. The method of embodiment 38, wherein said threshold is between 40 and 50 ng / mL. The method of embodiment 39, wherein said threshold is about 40 ng / mL. The method of embodiment 35, wherein said dose is between 2 to 20 mg / kg bodyweight. The method of embodiment 41, wherein said dose is between 3 to 15 mg / kg bodyweight. The method of embodiment 42, wherein said dose is between 5 to 12 mg / kg bodyweight. The method of embodiment 43, wherein said dose is between 7 to 10 mg / kg bodyweight. The method of embodiment 44, wherein said dose is about 10 mg / kg bodyweight. A method for the treatment of shock in an extreme-critically ill patient having shock, comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof according to any of embodiments 1 to 34; wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold of DPP3 concentration; wherein said threshold is an x-fold of the median level of DPP3 concentration in a healthy population, and wherein said threshold of DPP3 concentration is in the range between the 1.4-fold and 8.3-fold of the median of the DPP3 concentration in a healthy population; and wherein said anti-DPP3 antibody is administered at a dose between 1 to 30 mg / kg bodyweight. The method of embodiment 46, wherein said threshold of DPP3 concentration is in the range between 1.7-fold and 6.9-fold of the median of the DPP3 concentration in a healthy population. The method of embodiment 47, wherein said threshold of DPP3 concentration is in the range between 2.1-fold and 5.5-fold of the median ofthe DPP3 concentration in a healthy population. The method of embodiment 48, wherein said threshold of DPP3 concentration is in the range between 2.4 and 4. 1-fold of the median ofthe DPP3 concentration in a healthy population. The method of embodiment 49, wherein said threshold of DPP3 concentration is in the range between 2.8 and 3.4-fold of the median ofthe DPP3 concentration in a healthy population. The method of embodiment 50, wherein said threshold of DPP3 concentration is in the range between is about is the 2.8-fold of the median of the DPP3 concentration in a healthy population. The method of embodiment 46, wherein said dose is between 2 to 20 mg / kg bodyweight. The method of embodiment 52, wherein said dose is between 3 to 15 mg / kg bodyweight. The method of embodiment 53, wherein said dose is between 5 to 12 mg / kg bodyweight. The method of embodiment 54, wherein said dose is between 7 to 10 mg / kg bodyweight. The method of embodiment 55, wherein said dose is about 10 mg / kg bodyweight. A method for the treatment of shock in an extreme-critically ill patient having shock, comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof according to any of embodiments 1 to 34, wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold of DPP3 concentration, wherein said threshold of DPP3 concentration is 120 ng / ml or less, and wherein said anti-DPP3 antibody is administered at a dose between 1 to 30 mg / kg bodyweight. The method of embodiment 57, wherein said threshold of DPP3 concentration is 100 ng / mL or less. The method of embodiment 58, wherein said threshold of DPP3 concentration is 80 ng / mL or less. The method of embodiment 59, wherein said threshold of DPP3 concentration is 60 ng / ml or less. The method of embodiment 60, wherein said threshold of DPP3 concentration is 50 ng / ml or less. The method of embodiment 61, wherein said threshold of DPP3 concentration is about 40 ng / mL The method of embodiment 57, wherein said dose is between 2 to 20 mg / kg bodyweight. The method of embodiment 63, wherein said dose is between 3 to 15 mg / kg bodyweight. The method of embodiment 64, wherein said dose is between 5 to 12 mg / kg bodyweight. The method of embodiment 65, wherein said dose is between 7 to 10 mg / kg bodyweight. The method of embodiment 66, wherein said dose is about 10 mg / kg bodyweight. The method according to any of embodiments 1 to 67, wherein the shock is selected from the group consisting of hypovolemic shock, cardiogenic shock, obstructive shock, or distributive shock. The method according to any of embodiments 1 to 68, wherein the shock is cardiogenic shock. The method according to any of embodiments 1 to 69, wherein the distributive shock is septic shock. The method according to any of embodiments 1 to 72, wherein the shock is septic shock. Anti-DPP3 antibody thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiments 1 to 14, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody comprises the following sequences:

[0449] SEQ ID No. 34

[0450] QITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEWLAHIWWNDN KSYNPALKSRLTITRDTSKNQVVLTMTNMDPVDTGTYYCARNYSYDYWGQGTLVTV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP QVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF FLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG and / or

[0451] SEQ ID No. 35

[0452] DIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLYWYLQKPGQSPQLLIYKVSNRF SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPWTFGGGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC or alternatively sequences that are > 80% identical, preferably > 90% identical, preferably > 95% identical, preferably > 98% identical, preferably > 99% identical thereto, wherein the heavy chain comprises at least one CDR selected from the group comprising SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or

[0453] SEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR selected from the group comprising: SEQ ID No. 10: RSLVHSIGSTY, CDR2: KVS and / or

[0454] SEQ ID No. 11: SQSTHVPWT, or a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences, provided that the binding affinity to DPP3 is diminished by not more than 20%. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiments 1 to 14 and 72, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3antibody is administered at a dose between 1 to 30 mg / kg bodyweight, preferably 2 to 20 mg / kg bodyweight, preferably 3 to 15 mg / kg bodyweight, preferably 5 to 12 mg / kg body weight, preferably 7 to 10 mg / kg bodyweight and most preferred 10 mg / kg bodyweight. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiments 1 to 14 and 72 to 73, wherein an anti-DPP3antibody is used and wherein said anti-DPP3 is administered for 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiments 1 to 14 and 72 to 74, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody is administered at a dose of 10 mg / kg bodyweight and administered as infusion over 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 11 to 14 and 72 to 75, wherein the heavy chain comprises the CD Rs selected from the group comprising:

[0455] SEQ ID No. 7: GFSLSTSGMS,

[0456] SEQ ID No. 8: IWWNDNK and / or

[0457] SEQ ID No. 9: ARNYSYDY or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, according to embodiment 11 to 14 and 72 to 76, wherein the light chain comprises the CD Rs selected from the group comprising:

[0458] SEQ ID No. 10: RSLVHSIGSTY,

[0459] CDR2: KVS and / or

[0460] SEQ ID No. 11: SQSTHVPWT or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 11 to 14 and 72 to 77, wherein the heavy chain comprises the CD Rs selected from the group comprising:

[0461] SEQ ID No. 7: GFSLSTSGMS,

[0462] SEQ ID No. 8: IWWNDNK and / or

[0463] SEQ ID No. 9: ARNYSYDY. and wherein the light chain comprises the CDRs selected from the group comprising: SEQ ID No. 10: RSLVHSIGSTY,

[0464] CDR2: KVS and / or

[0465] SEQ ID No. 11: SQSTHVPWT or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiments 11 to 14 and 72 to 78, wherein said antibody or fragment thereof comprises a heavy chain Fab Fragment comprising SEQ ID NO: 38. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiments 11 to 14 and 72 to 79, wherein said antibody or fragment thereof comprises a light chain Fab Fragment comprising SEQ ID NO: 35. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 11 to 14 and 72 to 80, wherein said antibody or fragment thereof comprises a heavy chain Fab Fragment comprising SEQ ID NO: 138 and comprises a light chain Fab Fragment comprising SEQ ID NO: 35. It is apparent that, as in other related embodiments, “SEQ ID NO: 138” is meant to relate to SEQ ID NO: 38. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 11 to 14 and 72 to 81, wherein said antibody or fragment thereof comprises a heavy chain variable region portion of SEQ ID NO: 38. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 11 to 14 and 72 to 82, wherein said antibody or fragment thereof comprises a light chain variable region portion of SEQ ID NO: 35. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 11 to 14 and 72 to 83, wherein said antibody or fragment thereof comprises a heavy chain variable region portion of SEQ ID NO: 38 and comprises a light chain variable region portion of SEQ ID NO: 35. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 11 to 14 and 72 to 84, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a ScFv and wherein the ScFv comprises a heavy chain variable region portion of SEQ ID NO: 38 and comprises a light chain variable region portion of SEQ ID NO: 35 portion of SEQ ID NO: 35 and wherein the ScFv further comprises a linker connecting the heavy chain variable region portion of SEQ ID NO: 38 and the light chain variable region portion of SEQ ID NO: 35. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 11 to 14 and 72 to 85, wherein said antibody or fragment thereof comprises a heavy chain Fab region comprising SEQ ID NO: 38 and comprises a light chain variable region comprising SEQ ID NO: 35. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 11 to 14 and 72 to 86, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a ScFv and wherein the ScFv comprises a heavy chain variable region comprising SEQ ID NO: 38 and comprises a light chain variable region comprising SEQ ID NO: 35. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 11 to 14 and 72 to 87, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a Fab fragment and wherein the Fab fragment comprises a heavy chain Fab region comprising SEQ ID NO: 38 and comprises a light chain variable region comprising SEQ ID NO: 35. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 11 to 14 and 72 to 88, wherein said antibody or fragment thereof comprises a heavy chain variable region comprising SEQ ID NO: 5. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to any of embodiment 11 to 14 and 72 to 89, wherein said antibody or fragment thereof comprises a light chain variable region comprising SEQ ID NO: 6. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to embodiment 11 to 14 and 72 to 90, wherein said antibody or fragment thereof comprises a heavy chain variable region comprising SEQ ID NO: 5 and comprises a light chain variable region comprising SEQ ID NO: 6. A method for the treatment of shock in an extreme-critically ill patient having shock, comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof according to any of embodiments 1 to 14 and 72 to 91; wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold of DPP3 concentration; wherein said threshold is between 20 and 120 ng / mL; and wherein said anti-DPP3 antibody is administered at a dose between 1 and 30 mg / kg bodyweight. The method of embodiment 92, wherein said threshold is between 25 and 100 ng / mL. The method of embodiment 93, wherein said threshold is between 30 and 80 ng / mL. The method of embodiment 94, wherein said threshold is between 35 and 60 ng / mL. The method of embodiment 95, wherein said threshold is between 40 and 50 ng / mL. The method of embodiment 96, wherein said threshold is about 40 ng / mL. The method of embodiment 92, wherein said dose is between 2 to 20 mg / kg bodyweight. The method of embodiment 98, wherein said dose is between 3 to 15 mg / kg bodyweight. The method of embodiment 99, wherein said dose is between 5 to 12 mg / kg bodyweight. The method of embodiment 100, wherein said dose is between 7 to 10 mg / kg bodyweight. The method of embodiment 101, wherein said dose is about 10 mg / kg bodyweight. A method for the treatment of shock in an extreme-critically ill patient having shock, comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof according to any of embodiments 1 to 14 and 72 to 92; wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold of DPP3 concentration; wherein said threshold is an x-fold of the median level of DPP3 concentration in a healthy population, and wherein said threshold of DPP3 concentration is in the range between the 1.4-fold and 8.3-fold of the median of the DPP3 concentration in a healthy population; and wherein said anti-DPP3 antibody is administered at a dose between 1 to 30 mg / kg bodyweight. The method of embodiment 103, wherein said threshold of DPP3 concentration is in the range between 1.7-fold and 6.9-fold of the median of the DPP3 concentration in a healthy population. The method of embodiment 104, wherein said threshold of DPP3 concentration is in the range between 2.1-fold and 5.5-fold of the median ofthe DPP3 concentration in a healthy population. The method of embodiment 105, wherein said threshold of DPP3 concentration is in the range between 2.4 and 4. 1-fold of the median ofthe DPP3 concentration in a healthy population. The method of embodiment 106, wherein said threshold of DPP3 concentration is in the range between 2.8 and 3.4-fold of the median ofthe DPP3 concentration in a healthy population. The method of embodiment 107, wherein said threshold of DPP3 concentration is in the range between is about is the 2.8-fold of the median of the DPP3 concentration in a healthy population. The method of embodiment 103, wherein said dose is between 2 to 20 mg / kg bodyweight. The method of embodiment 104, wherein said dose is between 3 to 15 mg / kg bodyweight. The method of embodiment 105, wherein said dose is between 5 to 12 mg / kg bodyweight. The method of embodiment 106, wherein said dose is between 7 to 10 mg / kg bodyweight. The method of embodiment 107, wherein said dose is about 10 mg / kg bodyweight. A method for the treatment of shock in an extreme-critically ill patient having shock, comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof according to any of embodiments 1 to 14 and 72 to 92, wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold of DPP3 concentration, wherein said threshold of DPP3 concentration is 120 ng / ml or less, and wherein said anti-DPP3 antibody is administered at a dose between 1 to 30 mg / kg bodyweight. The method of embodiment 114, wherein said threshold of DPP3 concentration is 100 ng / mL or less. The method of embodiment 115, wherein said threshold of DPP3 concentration is 80 ng / mL or less. The method of embodiment 116, wherein said threshold of DPP3 concentration is 60 ng / ml or less. The method of embodiment 117, wherein said threshold of DPP3 concentration is 50 ng / ml or less. The method of embodiment 117, wherein said threshold of DPP3 concentration is about 40 ng / mL The method of embodiment 114, wherein said dose is between 2 to 20 mg / kg bodyweight. The method of embodiment 121, wherein said dose is between 3 to 15 mg / kg bodyweight. The method of embodiment 122, wherein said dose is between 5 to 12 mg / kg bodyweight. The method of embodiment 123, wherein said dose is between 7 to 10 mg / kg bodyweight. The method of embodiment 124, wherein said dose is about 10 mg / kg bodyweight. The method according to any of embodiments 1 to 14 and 72 to 124, wherein the shock is selected from the group consisting of hypovolemic shock, cardiogenic shock, obstructive shock, or distributive shock. The method according to any of embodiments 1 to 14 and 72 to 125, wherein the shock is cardiogenic shock. The method according to any of embodiments 1 to 14 and 72 to 126, wherein the distributive shock is septic shock. The method according to any of embodiments 1 to 14 and 72 to 127, wherein the shock is septic shock. In addition, the following consecutively numbered embodiments provide further specific aspects of the invention:

[0466] 72b Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, wherein said shock patient has acute kidney injury (AKI).

[0467] 73b Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiment 72b, wherein said critically ill patient with shock is under renal replacement therapy.

[0468] 74b Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b and 73b, wherein said shock is selected from the group comprising shock due to hypovolemia, cardiogenic shock, obstructive shock and distributive shock.

[0469] 75b Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 74b, wherein said patient has combined cardiogenic and septic shock.

[0470] 76b Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 75b, wherein said shock is refractory shock.

[0471] 77b Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 76b, wherein an anti-DPP3 antibody is used and wherein the heavy chain comprises at least one CDR, preferably at least three CD Rs selected from the group comprising:

[0472] SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or SEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR, preferably at least three CDRs selected from the group comprising:

[0473] SEQ ID No. 10: RSLVHSIGSTY, CDR2: KVS and / or SEQ ID No. 11: SQSTHVPWT.

[0474] 78b Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 77b, wherein said anti-DPP3 antibody comprises the following sequences:

[0475] SEQ ID No. 12 MDPKGSLSWRILLFLSLAFELSYGQITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQ PPGKALEWLAHIWWNDNKSYNPALKSRLTITRDTSKNQVVLTMTNMDPVDTGTYYCARNY SYDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQS SGLYSLS S VVTVPS S SLGTQTYICNVNHKP SNTKVDKKVEPKSCDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG and / or

[0476] SEQ ID No. 13

[0477] METDTLLLWVLLLWVPGSTGDIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLYWYLQ KPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPWTFGGG TKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC or a sequence that is > 80% identical to it, > 90% identical to it, > 95% identical to it, preferably > 98%, preferably > 99%.

[0478] 79b Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 78b, wherein said anti-DPP3 antibody is administered at a dose from 1 to 30 mg / kg bodyweight, preferably 2 to 20 mg / kg bodyweight, preferably 3 to 15 mg / kg bodyweight, preferably 5 to 12 mg / kg body weight, preferably 7 to 10 mg / kg bodyweight, more preferably 10 mg / kg bodyweight.

[0479] 80b Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 79b, wherein said anti-DPP3 is administered over the course of 1 to 3 hours, preferably 1.5 to 2.5 hours, more preferably 2 hours and / or wherein said anti-DPP3 antibody is administered at a dose of 10 mg / kg bodyweight as infusion over the course of 1 to 3 hours, preferably 1.5 to 2.5 hours, more preferably 2 hours.

[0480] 81b Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 80b, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody comprises the following sequences:

[0481] SEQ ID No. 34

[0482] QITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEWLAHIWWNDNKSYNP ALKSRLTITRDTSKNQVVLTMTNMDPVDTGTYYCARNYSYDYWGQGTLVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV PSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPG and / or

[0483] SEQ ID No. 35

[0484] DIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLYWYLQKPGQSPQLLIYKVSNRFSGVPD RFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLS SP VTKSFNRGEC or alternatively sequences that are > 80% identical, preferably > 90% identical, preferably > 95% identical, preferably > 98% identical, preferably > 99% identical thereto, wherein the heavy chain comprises at least one CDR selected from the group comprising SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or SEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR selected from the group comprising: SEQ ID No. 10: RSLVHSIGSTY, CDR2: KVS and / or SEQ ID No. 11: SQSTHVPWT, or a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences, provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0485] 82b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 81b, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody is administered at a dose between 1 to 30 mg / kg bodyweight, preferably 2 to 20 mg / kg bodyweight, preferably 3 to 15 mg / kg bodyweight, preferably 5 to 12 mg / kg body weight, preferably 7 to 10 mg / kg bodyweight and most preferred 10 mg / kg bodyweight.

[0486] 83b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 82b, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 is administered for 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours.

[0487] 84b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 83b, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody is administered at a dose of 10 mg / kg bodyweight and administered as infusion over 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours. 85b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 84b, wherein the heavy chain comprises the CDRs selected from the group comprising:

[0488] SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or SEQ ID No. 9: ARNYSYDY or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0489] 86b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 85b, wherein the light chain comprises the CDRs selected from the group comprising:

[0490] SEQ ID No. 10: RSLVHSIGSTY, CDR2: KVS and / or SEQ ID No. 11: SQSTHVPWT or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0491] 87b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 86b, wherein the heavy chain comprises the CDRs selected from the group comprising:

[0492] SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or SEQ ID No. 9: ARNYSYDY. and wherein the light chain comprises the CDRs selected from the group comprising:

[0493] SEQ ID No. 10: RSLVHSIGSTY, CDR2: KVS and / or SEQ ID No. 11: SQSTHVPWT or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0494] 88b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 87b, wherein said antibody or fragment thereof comprises a heavy chain Fab Fragment comprising SEQ ID NO: 38.

[0495] 89b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 88b, wherein said antibody or fragment thereof comprises a light chain Fab Fragment comprising SEQ ID NO: 35. 90b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 89b, wherein said antibody or fragment thereof comprises a heavy chain Fab Fragment comprising SEQ ID NO: 138 and comprises a light chain Fab Fragment comprising SEQ ID NO: 35.

[0496] 91b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 90b, wherein said antibody or fragment thereof comprises a heavy chain variable region portion of SEQ ID NO: 38.

[0497] 92b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 91b, wherein said antibody or fragment thereof comprises a light chain variable region portion of SEQ ID NO: 35.

[0498] 93b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 92b, wherein said antibody or fragment thereof comprises a heavy chain variable region portion of SEQ ID NO: 38 and comprises a light chain variable region portion of SEQ ID NO: 35.

[0499] 94b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 93b, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a ScFv and wherein the ScFv comprises a heavy chain variable region portion of SEQ ID NO: 38 and comprises a light chain variable region portion of SEQ ID NO: 35 portion of SEQ ID NO: 35 and wherein the ScFv further comprises a linker connecting the heavy chain variable region portion of SEQ ID NO: 38 and the light chain variable region portion of SEQ ID NO: 35.

[0500] 95b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 94b, wherein said antibody or fragment thereof comprises a heavy chain Fab region comprising SEQ ID NO: 38 and comprises a light chain variable region comprising SEQ ID NO: 35.

[0501] 96b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 95b, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a ScFv and wherein the ScFv comprises a heavy chain variable region comprising SEQ ID NO: 38 and comprises a light chain variable region comprising SEQ ID NO: 35.

[0502] 97b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 96b, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a Fab fragment and wherein the Fab fragment comprises a heavy chain Fab region comprising SEQ ID NO: 38 and comprises a light chain variable region comprising SEQ ID NO: 35.

[0503] 98b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 97b, wherein said antibody or fragment thereof comprises a heavy chain variable region comprising SEQ ID NO: 5.

[0504] 99b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 98b, wherein said antibody or fragment thereof comprises a light chain variable region comprising SEQ ID NO: 6.

[0505] 100b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 99b, wherein said antibody or fragment thereof comprises a heavy chain variable region comprising SEQ ID NO: 5 and comprises a light chain variable region comprising SEQ ID NO: 6.

[0506] 101b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 100b; wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold of DPP3 concentration; wherein said threshold is between 20 and 120 ng / mL, particularly between 25 and 100 ng / mL, more particularly between 30 and 80 ng / mL, even more particularly between 35 and 60 ng / mL, even more particularly between 40 and 50 ng / mL, even more particularly said threshold is about 40 ng / mL; and wherein said anti-DPP3 antibody is administered at a dose between 1 and 30 mg / kg bodyweight, particularly between 2 to 20 mg / kg bodyweight, more particularly between 3 to 15 mg / kg bodyweight, even more particularly between 5 to 12 mg / kg body weight, even more particularly between 7 to 10 mg / kg bodyweight, even more particularly said dose is about 10 mg / kg bodyweight. 102b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 100b; wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold of DPP3 concentration; wherein said threshold is an x-fold of the median level of DPP3 concentration in a healthy population, and wherein said threshold of DPP3 concentration is in the range between the 1.4-fold and 8.3-fold, particularly in the range between 1.7-fold and 6.9-fold, more particularly in the range between 2. 1-fold and 5.5-fold, even more particularly in the range between 2.4 and 4.1-fold, even more particularly in the range between 2.8 and 3.4-fold, even more particularly about the 2.8-fold, of the median of the DPP3 concentration in a healthy population; and wherein said anti-DPP3 antibody is administered at a dose between 1 to 30 mg / kg bodyweight, particularly between 2 to 20 mg / kg bodyweight, more particularly between 3 to 15 mg / kg bodyweight, even more particularly between 5 to 12 mg / kg body weight, even more particularly between 7 to 10 mg / kg bodyweight, even more particularly said dose is about 10 mg / kg bodyweight.

[0507] 103b. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to embodiments 72b to 100b; wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold of DPP3 concentration, wherein said threshold of DPP3 concentration is 120 ng / ml or less, particularly 100 ng / mL or less, more particularly 80 ng / mL or less, even more particularly 60 ng / ml or less, even more particularly 50 ng / ml or less, even more particularly about 40 ng / mL, and wherein said anti-DPP3 antibody is administered at a dose between 1 to 30 mg / kg bodyweight, particularly between 2 to 20 mg / kg bodyweight, more particularly between 3 to 15 mg / kg bodyweight, even more particularly between 5 to 12 mg / kg body weight, even more particularly between 7 to 10 mg / kg bodyweight, even more particularly said dose is about 10 mg / kg bodyweight.

[0508] In addition, the following consecutively numbered embodiments provide further specific aspects of the invention:

[0509] 1c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, wherein said patient is an extreme-critically ill patient, wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and wherein the level of DPP3 is determined by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3.

[0510] 2c. Method comprising determining the level of DPP3 in a sample of a bodily fluid of a patient having shock by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and, if said level of DPP3 in said sample is above a threshold, categorizing the patient as being extreme-critically ill and as requiring administration of an anti-DPP3 antibody or fragment thereof for the treatment of said shock.

[0511] 3c. A method for the treatment of shock in an extreme-critically ill patient having shock, comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof, wherein said extreme-critically ill patient is characterized by a level of DPP3 in a sample of a bodily fluid of said patient above a threshold, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and wherein the level of DPP3 is determined by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP 3.

[0512] 4c. A method for the treatment of shock in an extreme-critically ill patient having shock, comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof, wherein the level of DPP3 determined in a sample of a bodily fluid of said patient by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3 is above a threshold, and wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample.

[0513] 5c. A method comprising determining the level of DPP3 in a sample of a bodily fluid of an extreme- critically ill patient having shock by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and, if said level of DPP3 in said sample is above a threshold, administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof.

[0514] 6c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, wherein said shock patient has acute kidney injury (AKI), wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and wherein the level of DPP3 is determined by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3.

[0515] 7c. A Method comprising determining the level of DPP3 in a sample of a bodily fluid of a patient with shock and having acute kidney injury (AKI) by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP 3, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and, if said level of DPP3 in said sample is above a threshold, categorizing the patient as being critically ill and as requiring administration of an anti-DPP3 antibody or fragment thereof for use in therapy of said patient and for improvement or stabilization of kidney function in said patient.

[0516] 8c. A method for the treatment of shock in a critically ill patient with shock and having acute kidney injury (AKI), comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof, wherein said critically ill patient is characterized by a level of DPP3 in a sample of a bodily fluid of said patient above a threshold, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and wherein the level of DPP3 is determined by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3.

[0517] 9c. A method for the treatment of shock in a critically ill patient with shock and having acute kidney injury (AKI), comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof, wherein the level of DPP3 determined in a sample of a bodily fluid of said patient by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3 is above a threshold, and wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample.

[0518] 10c. A method comprising determining the level of DPP3 in a sample of a bodily fluid of a critically ill patient with shock and having acute kidney injury (AKI) by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and, if said level of DPP3 in said sample is above a threshold, administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof.

[0519] 11c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 10c, wherein said shock is selected from the group comprising hypovolemic shock, cardiogenic shock, obstructive shock and distributive shock.

[0520] 12c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 11c, wherein said shock patient has combined cardiogenic and septic shock. 13c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 12c, wherein said shock is refractory shock.

[0521] 14c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 13c, wherein said extreme-critically ill patient has a 24-hour probability of mortality of at least 70%, preferably of at least 80%, more preferably of at least 90%, most preferably of at least 95% and / or has organ dysfunction requiring mechanical or pharmacological support of at least two organs.

[0522] 15c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 14c, wherein said organ dysfunction is defined as cardiovascular dysfunction, respiratory dysfunction (lung), kidney dysfunction, liver dysfunction, central nervous system dysfunction or dysfunction of the coagulation system.

[0523] 16c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 14c or 15c, wherein said organ support is selected from kidney support, respiratory (lung) support, cardiovascular support and / or liver support.

[0524] 17c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 16c, wherein said level of DPP3 is the DPP3 concentration and wherein said threshold of DPP 3 concentration is between 20 and 120 ng / mL, more preferred between 25 and 100 ng / mL, even more preferred between 30 and 80 ng / mL, even more preferred between 35 and 60 ng / mL, even more preferred between 40 and 50 ng / ml most preferred said threshold is 40 ng / mL.

[0525] 18c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 16c, wherein said level of DPP3 is the DPP3 the enzymatic activity of cDPP3 and wherein said threshold of enzymatic activity of DPP3 is between 1.76 and 9.68 pmol / (min*L), more preferred between 2.15 and 8.09 pmol / (min*L), even more preferred between 2.55 and 6.51 pmol / (min*L), even more preferred between 2.94 and 4.93 pmol / (min*L), even more preferred between 3.34 and 4. 13 pmol / (min*L), most preferred said threshold is 3.34 pmol / (min*L).

[0526] 19c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 18c, wherein said bodily fluid is selected from the group comprising whole blood, serum and plasma.

[0527] 20c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 19c, wherein the patient is characterized by a level of lactate of 2 mmol / 1 or higher and / or a level of IL-6 of 5.9 pg / ml or higher and / or a level of CRP of 1 mg / dL or higher and / or a level of PCT of 0.5 ng / ml or higher and / or a level ofbio-ADM of 70 pg / ml or higher and / or serum creatinine of 1.18 mg / dL (for men) and 1.02 mg / dL (for women) and / or a supranormal level of NT-proBNP in a sample of bodily fluid of said patient.

[0528] 21c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 20c, wherein an anti-DPP3antibody is used and wherein the heavy chain comprises at least one CDR, preferably at least three CD Rs selected from the group comprising:

[0529] SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or SEQ ID No. 9: ARNYSYDY. and wherein the light chain comprises at least one CDR, preferably at least three CD Rs selected from the group comprising:

[0530] SEQ ID No. 10: RSLVHSIGSTY, CDR2: KVS and / or SEQ ID No. 11: SQSTHVPWT or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0531] 22c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody comprises the following sequences:

[0532] SEQ ID No. 12 MDPKGSLSWRILLFLSLAFELSYGQITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQ PPGKALEWLAHIWWNDNKSYNPALKSRLTITRDTSKNQVVLTMTNMDPVDTGTYYCARNY SYDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQS SGLYSLS S VVTVPS S SLGTQTYICNVNHKP SNTKVDKKVEPKSCDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG and / or

[0533] SEQ ID No. 13

[0534] METDTLLLWVLLLWVPGSTGDIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLYWYLQ KPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPWTFGGG TKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC or alternatively sequences that are > 80% identical, preferably > 90% identical, preferably > 95% identical, preferably > 98% identical, preferably > 99% identical thereto, wherein the heavy chain comprises at least one CDR selected from the group comprising SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or SEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR selected from the group comprising: SEQ ID No. 10: RSLVHSIGSTY, CDR2: KVS and / or SEQ ID No. 11: SQSTHVPWT, or a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0535] 23c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 22c, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody is administered at a dose between 1 to 30 mg / kg bodyweight, preferably 2 to 20 mg / kg bodyweight, preferably 3 to 15 mg / kg bodyweight, preferably 5 to 12 mg / kg body weight, preferably 7 to 10 mg / kg bodyweight and most preferred 10 mg / kg bodyweight.

[0536] 24c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 23c, wherein an anti-DPP3antibody is used and wherein said anti-DPP3 is administered for 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours. 25c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 24c, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody is administered at a dose of 10 mg / kg bodyweight and administered as infusion over 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours.

[0537] 26c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 25c, wherein the heavy chain comprises the CDRs selected from the group comprising:

[0538] SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or SEQ ID No. 9: ARNYSYDY or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0539] 27c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 26c, wherein the light chain comprises the CDRs selected from the group comprising:

[0540] SEQ ID No. 10: RSLVHSIGSTY, CDR2: KVS and / or SEQ ID No. 11: SQSTHVPWT or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0541] 28c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 27c, wherein the heavy chain comprises the CDRs selected from the group comprising:

[0542] SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or SEQ ID No. 9: ARNYSYDY. and wherein the light chain comprises the CDRs selected from the group comprising:

[0543] SEQ ID No. 10: RSLVHSIGSTY, CDR2: KVS and / or SEQ ID No. 11: SQSTHVPWT or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%. 29c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 28c, wherein said antibody or fragment thereof comprises a heavy chain Fab Fragment comprising SEQ ID NO: 16.

[0544] 30c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 29c, wherein said antibody or fragment thereof comprises a light chain Fab Fragment comprising SEQ ID NO: 13.

[0545] 31c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 30c, wherein said antibody or fragment thereof comprises a heavy chain Fab Fragment comprising SEQ ID NO: 16 and comprises a light chain Fab Fragment comprising SEQ ID NO: 13.

[0546] 32c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 31c, wherein said antibody or fragment thereof comprises a heavy chain variable region portion of SEQ ID NO: 16.

[0547] 33c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 32c, wherein said antibody or fragment thereof comprises a light chain variable region portion of SEQ ID NO: 13.

[0548] 34c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 33c, wherein said antibody or fragment thereof comprises a heavy chain variable region portion of SEQ ID NO: 16 and comprises a light chain variable region portion of SEQ ID NO: 13.

[0549] 35c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 34c, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a ScFv and wherein the ScFv comprises a heavy chain variable region portion of SEQ ID NO: 16 and comprises a light chain variable region portion of SEQ ID NO: 13 portion of SEQ ID NO: 13 and wherein the ScFv further comprises a linker connecting the heavy chain variable region portion of SEQ ID NO: 16 and the light chain variable region portion of SEQ ID NO: 13.

[0550] 36c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 35c, wherein said antibody or fragment thereof comprises a heavy chain Fab region comprising SEQ ID NO: 16 and comprises a light chain variable region comprising SEQ ID NO: 13.

[0551] 37c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 36c, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a ScFv and wherein the ScFv comprises a heavy chain variable region comprising SEQ ID NO: 16 and comprises a light chain variable region comprising SEQ ID NO: 13.

[0552] 38c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 37c, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a Fab fragment and wherein the Fab fragment comprises a heavy chain Fab region comprising SEQ ID NO: 16 and comprises a light chain variable region comprising SEQ ID NO: 13.

[0553] 39c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 38c, wherein said antibody or fragment thereof comprises a heavy chain variable region comprising SEQ ID NO: 5.

[0554] 40c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 39c, wherein said antibody or fragment thereof comprises a light chain variable region comprising SEQ ID NO: 6. 41c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 40c, wherein said antibody or fragment thereof comprises a heavy chain variable region comprising SEQ ID NO: 5 and comprises a light chain variable region comprising SEQ ID NO: 6.

[0555] 42c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody comprises the following sequences: SEQ ID No. 34

[0556] QITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEWLAHIWWNDNKSYNP ALKSRLTITRDTSKNQVVLTMTNMDPVDTGTYYCARNYSYDYWGQGTLVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV PSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPG and / or

[0557] SEQ ID No. 35

[0558] DIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLYWYLQKPGQSPQLLIYKVSNRFSGVPD RFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLS SP VTKSFNRGEC or alternatively sequences that are > 80% identical, preferably > 90% identical, preferably > 95% identical, preferably > 98% identical, preferably > 99% identical thereto, wherein the heavy chain comprises at least one CDR selected from the group comprising SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or SEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR selected from the group comprising: SEQ ID No. 10: RSLVHSIGSTY, SDR2: KVS and / or SEQ ID No. 11: SQSTHVPWT, or a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%. 43c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c or 42c, wherein an anti-DPP3 antibody is used and wherein said anti- DPP3antibody is administered at a dose between 1 to 30 mg / kg bodyweight, preferably 2 to 20 mg / kg bodyweight, preferably 3 to 15 mg / kg bodyweight, preferably 5 to 12 mg / kg body weight, preferably 7 to 10 mg / kg bodyweight and most preferred 10 mg / kg bodyweight.

[0559] 44c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, 42c or 43c, wherein an anti-DPP3antibody is used and wherein said anti-DPP3 is administered for 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours.

[0560] 45c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 44c, wherein an anti-DPP3 antibody is used and wherein said anti- DPP3 antibody is administered at a dose of 10 mg / kg bodyweight and administered as infusion over 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours.

[0561] 46c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 45c, wherein the heavy chain comprises the CDRs selected from the group comprising:

[0562] SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or SEQ ID No. 9: ARNYSYDY or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0563] 47c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 46c, wherein the light chain comprises the CDRs selected from the group comprising:

[0564] SEQ ID No. 10: RSLVHSIGSTY, CDR2: KVS and / or SEQ ID No. 11: SQSTHVPWT or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0565] 48c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 47c, wherein the heavy chain comprises the CDRs selected from the group comprising:

[0566] SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / or SEQ ID No. 9: ARNYSYDY. and wherein the light chain comprises the CDRs selected from the group comprising:

[0567] SEQ ID No. 10: RSLVHSIGSTY, CDR2: KVS and / or SEQ ID No. 11: SQSTHVPWT or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

[0568] 49c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 48c, wherein said antibody or fragment thereof comprises a heavy chain Fab Fragment comprising SEQ ID NO: 38.

[0569] 50c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 49c, wherein said antibody or fragment thereof comprises a light chain Fab Fragment comprising SEQ ID NO: 35.

[0570] 51c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 50c, wherein said antibody or fragment thereof comprises a heavy chain Fab Fragment comprising SEQ ID NO: 138 and comprises a light chain Fab Fragment comprising SEQ ID NO: 35.

[0571] 52c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 51c, wherein said antibody or fragment thereof comprises a heavy chain variable region portion of SEQ ID NO: 38. 53c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 52c, wherein said antibody or fragment thereof comprises a light chain variable region portion of SEQ ID NO: 35.

[0572] 54c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 53c, wherein said antibody or fragment thereof comprises a heavy chain variable region portion of SEQ ID NO: 38 and comprises a light chain variable region portion of SEQ ID NO: 35.

[0573] 55c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 54c, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a ScFv and wherein the ScFv comprises a heavy chain variable region portion of SEQ ID NO: 38 and comprises a light chain variable region portion of SEQ ID NO: 35 portion of SEQ ID NO: 35 and wherein the ScFv further comprises a linker connecting the heavy chain variable region portion of SEQ ID NO: 38 and the light chain variable region portion of SEQ ID NO: 35.

[0574] 56c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 55c, wherein said antibody or fragment thereof comprises a heavy chain Fab region comprising SEQ ID NO: 38 and comprises a light chain variable region comprising SEQ ID NO: 35.

[0575] 57c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 56c, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a ScFv and wherein the ScFv comprises a heavy chain variable region comprising SEQ ID NO: 38 and comprises a light chain variable region comprising SEQ ID NO: 35.

[0576] 58c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 57c, wherein said antibody or fragment thereof is a fragment, wherein the fragment is a Fab fragment and wherein the Fab fragment comprises a heavy chain Fab region comprising SEQ ID NO: 38 and comprises a light chain variable region comprising SEQ ID NO: 35.

[0577] 59c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 58c, wherein said antibody or fragment thereof comprises a heavy chain variable region comprising SEQ ID NO: 5.

[0578] 60c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 59c, wherein said antibody or fragment thereof comprises a light chain variable region comprising SEQ ID NO: 6.

[0579] 61c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 60c, wherein said antibody or fragment thereof comprises a heavy chain variable region comprising SEQ ID NO: 5 and comprises a light chain variable region comprising SEQ ID NO: 6.

[0580] 62c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 21c, or 42c to 61c, wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold of DPP3 concentration; wherein said threshold is an x-fold of the median level of DPP3 concentration in a healthy population, and wherein said threshold of DPP3 concentration is in the range between the 1.4-fold and 8.3-fold of the median of the DPP3 concentration in a healthy population; and wherein said anti-DPP3 antibody is administered at a dose between 1 to 30 mg / kg bodyweight.

[0581] 63c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to embodiment 62c, wherein said threshold of DPP3 concentration is in the range between 1.7-fold and 6.9-fold of the median of the DPP3 concentration in a healthy population, particularly in the range between 2.1 -fold and 5.5-fold of the median of the DPP3 concentration in a healthy population, more particularly in the range between 2.4 and 4.1-fold of the median of the DPP3 concentration in a healthy population, more particularly in the range between 2.8 and 3.4-fold of the median of the DPP3 concentration in a healthy population, more particularly about the 2.8 -fold of the median of the DPP3 concentration in a healthy population.

[0582] 64c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 63c, wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold of DPP3 concentration, wherein said threshold of DPP3 concentration is 120 ng / ml or less, and wherein said anti-DPP3 antibody is administered at a dose between 1 to 30 mg / kg bodyweight.

[0583] 65c. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient, or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, or method according to any one of embodiments 1c to 64c, wherein said threshold of DPP3 concentration is 100 ng / mL or less, particularly 80 ng / mL or less, more particularly 60 ng / ml or less, more particularly 50 ng / ml or less, more particularly about 40 ng / mL.

[0584] SEQUENCES

[0585] SEQ ID No. 1 - hDPP3 aa 1-737

[0586] MADTQYILPNDIGVSSLDCREAFRLLSPTERLYAYHLSRAAWYGGLAVLLQTSPEAPYI YALLSRLFRAQDPDQLRQHALAEGLTEEEYQAFLVYAAGVYSNMGNYKSFGDTKFV PNLPKEKLERVILGSEAAQQHPEEVRGLWQTCGELMFSLEPRLRHLGLGKEGITTYFS GNCTMEDAKLAQDFLDSQNLSAYNTRLFKEVDGEGKPYYEVRLASVLGSEPSLDSEV TSKLKSYEFRGSPFQVTRGDYAPILQKVVEQLEKAKAYAANSHQGQMLAQYIESFTQ GSIEAHKRGSRFWIQDKGPIVESYIGFIESYRDPFGSRGEFEGFVAVVNKAMSAKFERL VASAEQLLKELPWPPTFEKDKFLTPDFTSLDVLTFAGSGIPAGINIPNYDDLRQTEGFKN VSLGNVLAVAYATQREKLTFLEEDDKDLYILWKGPSFDVQVGLHELLGHGSGKLFVQ DEKGAFNFDQETVINPETGEQIQSWYRSGETWDSKFSTIASSYEECRAESVGLYLCLH PQVLEIFGFEGADAEDVIYVNWLNMVRAGLLALEFYTPEAFNWRQAHMQARFVILR VLLEAGEGLVTITPTTGSDGRPDARVRLDRSKIRSVGKPALERFLRRLQVLKSTGDVA GGRALYEGYATVTDAPPECFLTLRDTVLLRKESRKLIVQPNTRLEGSDVQLLEYEASA AGLIRSFSERFPEDGPELEEILTQLATADARFWKGPSEAPSGQA SEQ ID No. 2 - hDPP3 aa 474-493 (N-Cys) - immunization peptide with additional N- terminal Cystein

[0587] CETVINPETGEQIQSWYRSGE

[0588] SEQ ID No. 3 - hDPP3 aa 477-482 - epitope anti-DPP3 antibody

[0589] INPETG

[0590] SEQ ID No. 4 - hDPP3 aa 480-483

[0591] ETGE

[0592] SEQ ID No. 5 - variable region of murine anti-DPP3 antibody in heavy chain

[0593] QVTLKESGPGILQPSQTLSLTCSFSGFSLSTSGMSVGWIRQPSGKGLEWLAHIWWNDN

[0594] KSYNPALKSRLTISRDTSNNQVFLKIASVVTADTGTYFCARNYSYDYWGQGTTLTVSS

[0595] SEQ ID No. 6 - variable region of murine anti-DPP3 antibody in light chain

[0596] DVVVTQTPLSLSVSLGDPASISCRSSRSLVHSIGSTYLHWYLQKPGQSPKLLIYKVSNR

[0597] FSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYFCSQSTHVPWTFGGGTKLEIK

[0598] SEQ ID No. 7 - CDR1 of anti-DPP3 antibody in heavy chain

[0599] GFSLSTSGMS

[0600] SEQ ID No. 8 - CDR2 of anti-DPP3 antibody in heavy chain

[0601] IWWNDNK

[0602] SEQ ID No. 9 - CDR 3 of anti-DPP3 antibody in heavy chain

[0603] ARNYSYDY

[0604] SEQ ID No. 10 - CDR1 of anti-DPP3 antibody in light chain

[0605] RSLVHSIGSTY

[0606] CDR2 of anti-DPP3 antibody in light chain

[0607] KVS SEQ ID No. 11 - CDR3 of anti-DPP3 antibody in light chain

[0608] SQSTHVPWT

[0609] SEQ ID No. 12 - humanized anti-DPP3 antibody - heavy chain sequence (IgGlK backbone)

[0610] MDPKGSLSWRILLFLSLAFELSYGQITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSV GWIRQPPGKALEWLAHIWWNDNKSYNPALKSRLTITRDTSKNQVVLTMTNMDPVDT GTYYCARNYSYDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT KVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK VSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPG

[0611] SEQ ID No. 13 - humanized anti-DPP3 antibody - light chain sequence (IgGlK backbone)

[0612] METDTLLLWVLLLWVPGSTGDIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLY WYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQST HVPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC

[0613] SEQ ID No. 14 - human DPP3 amino acid 450-455

[0614] HELLGH

[0615] SEQ ID No. 15 - human DPP3 507-512

[0616] EECRAE

[0617] SEQ ID NO: 16 - humanized anti-DPP3 antibody - heavy chain sequence (IgGlK backbone) after treatment with Papain (Fab fragment)

[0618] MDPKGSLSWRILLFLSLAFELSYGQITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSV GWIRQPPGKALEWLAHIWWNDNKSYNPALKSRLTITRDTSKNQVVLTMTNMDPVDT GTYYCARNYSYDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT

[0619] KVDKKVEPKSCDKTH

[0620] SEQ ID NO: 17 - Peptide # 1 for Epitope mapping bio-AFNFDQETVINPETGEQIQSWYRS

[0621] SEQ ID NO: 18 - Peptide # 2 for Epitope mapping bio-AFNFDQETVINPETGEQIQ

[0622] SEQ ID NO: 19 - Peptide # 3 for Epitope mapping bio-AFNFDQETVINPETGEQI

[0623] SEQ ID NO: 20 - Peptide # 4 for Epitope mapping bio-AFNFDQETVINPETGEQ

[0624] SEQ ID NO: 21 - Peptide # 5 for Epitope mapping bio-AFNFDQETVINPETGE

[0625] SEQ ID NO: 22 - Peptide # 6 for Epitope mapping bio-AFNFDQETVINPETG

[0626] SEQ ID NO: 23 - Peptide # 7 for Epitope mapping bio-AFNFDQETVINPET

[0627] SEQ ID NO: 24 - Peptide # 8 for Epitope mapping bio-AFNFDQETVINPE

[0628] SEQ ID NO: 25 - Peptide # 9 for Epitope mapping bio-AFNFDQETVINP

[0629] SEQ ID NO: 26 - Peptide # 10 for Epitope mapping bio-AFNFDQETVIN

[0630] SEQ ID NO: 27 - Peptide # 11 for Epitope mapping ETGEQIQSWYK-bio

[0631] SEQ ID NO: 28 - Peptide # 12 for Epitope mapping

[0632] PETGEQIQSWYK-bio

[0633] SEQ ID NO: 29 - Peptide # 13 for Epitope mapping

[0634] NPETGEQIQSWYK-bio

[0635] SEQ ID NO: 30 - Peptide # 14 for Epitope mapping

[0636] INPETGEQIQSWYK-bio

[0637] SEQ ID NO: 31 - Peptide # 15 for Epitope mapping

[0638] VINPETGEQIQSWYK-bio

[0639] SEQ ID NO: 32 - Peptide # 16 for Epitope mapping

[0640] T VINPETGEQIQSWYK-bio

[0641] SEQ ID NO: 33 - Peptide # 17 for Epitope mapping

[0642] ETVINPETGEQIQSWYK-bio

[0643] SEQ ID NO: 34 - humanized anti-DPP3 antibody (PCZ) - heavy chain sequence (IgGlK backbone)

[0644] QITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEWLAHIWWNDN KSYNPALKSRLTITRDTSKNQVVLTMTNMDPVDTGTYYCARNYSYDYWGQGTLVTV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPE LLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK PREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL YSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0645] SEQ ID NO: 35 - humanized anti-DPP3 antibody (PCZ)- light chain sequence (IgGlK backbone) DIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLYWYLQKPGQSPQLLIYKVSNRF SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPWTFGGGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0646] SEQ ID NO: 36 - humanized anti-DPP3 antibody - heavy chain N-terminal signal peptide sequence

[0647] MDPKGSLSWRILLFLSLAFELSYG

[0648] SEQ ID NO: 37 - humanized anti-DPP3 antibody - light chain N-terminal signal peptide sequence

[0649] METDTLLLWVLLLWVPGSTG

[0650] SEQ ID NO: 38 - humanized anti-DPP3 antibody (PCZ) - heavy chain sequence (IgGlK backbone) after treatment with Papain (Fab fragment)

[0651] QITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEWLAHIWWNDN KSYNPALKSRLTITRDTSKNQVVLTMTNMDPVDTGTYYCARNYSYDYWGQGTLVTV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL

[0652] QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTH

[0653] SEQ ID NO: 39 - CDR1 of AK2555 heavy chain

[0654] GYYIH

[0655] SEQ ID No. 40 - CDR2 of AK2555 heavy chain

[0656] EINPSTGGTNYNQKFKG

[0657] SEQ ID No. 41 - CDR 3 of AK2555 heavy chain

[0658] SYYGIYSWFAY

[0659] SEQ ID No. 42 - CDR1 of AK2555 light chain

[0660] KASQDIKSYLS

[0661] SEQ ID No. 43 - CDR2 of AK2555 light chain

[0662] YATSLAD SEQ ID No. 44 - CDR3 of AK2555 light chain

[0663] LQHGESPRT

[0664] SEQ ID No. 45 - AK2555 - variable region of murine anti-DPP3 antibody in heavy chain sequence EVQLQQSGPELVKPGASVKISCKASGYSFTGYYIHWVKQSPENSLEWIGEINPSTGGTNYNQK FKGKATLTVDKSSSTAYMQLKSLTSEESAVYYCTRSYYGIYSWFAYWGQGTLVTVSA

[0665] SEQ ID No. 46 - AK2555 - variable region of murine anti-DPP3 antibody in light chain sequence DIKMTQSPSSMYASLGERVTITCKASQDIKSYLSWYQQKPWKSPKTLIYYATSLADGVPSRFSG SGSGQDYSLTISSLESDDTATYYCLQHGESPRTFGGGTKLEIK

[0666] Figure description

[0667] Figure 1: Inhibition curve of native DPP3 from blood cells with inhibitory murine anti-DPP3 antibody (cAK1967). Inhibition of DPP3 by a specific antibody is concentration dependent, with an IC50 at ~15 ng / ml when analysed against 15 ng / ml DPP3.

[0668] Figure 2: Association and dissociation curve of the murine anti-DPP3 antibody-DPP3 binding analysis using Octet. Murine anti-DPP3 antibody loaded biosensors were dipped into a dilution series of recombinant GST-tagged human DPP3 (100, 33.3, 11.1, and 3.7 nM) and association and dissociation monitored.

[0669] Figure 3: Western Blot of dilutions of blood cell lysate and detection of dilutions of blood cell lysate and recombinant human DPP3 with murine anti-hDPP3 antibody as primary antibody.

[0670] Figure 4 A and 4 B: Norepinephrine dosing before (“pre-dose”) and after (“24 h and 48 h post-dose”) treatment with PCZ. Shown are mean / SD values obtained from the three patients treated with PCZ. Figure 4 A shows the absolute dosages, Figure 4 B shows the dosages relative to the pre-dose dosages.

[0671] Figure 5: Blood lactate concentrations before (“pre-dose”) and after (“24 h and 48 h post-dose”) treatment with PCZ. Shown are mean / SD values obtained from the three patients treated with PCZ. Hyperlactatemia is defined as a lactate concentration above 2 mmol / L (indicated by the dotted line) (Vieira, Petrova, and Moura 2022).

[0672] Figure 6: Blood IL-6 concentrations before (“pre-dose”) and after (“24 h and 48 h post-dose”) treatment with PCZ. Shown are mean / SD values obtained from the three patients treated with PCZ. The upper limit of the healthy normal range is indicated by a dotted line. Figure 7 A and 7 B: Kidney function before (“pre-dose”) and after (“24 h and 48 h post-dose”) treatment with PCZ. Shown are mean / SD values obtained from the three patients treated with PCZ for blood creatinine concentrations (Figure 7 A) and the Glomerular Filtration Rate (GFR) calculated by the CKD- EPI formula (Figure 7 B). Limits of healthy normal ranges are indicated by dotted lines (between 0.55 and 1.02 mg / dL creatinine, and between 90 and 120 mL / min for the GFR).

[0673] Figure 8: Blood NT-proBNP concentrations before (“pre-dose”) and after (“24 h and 48 h post-dose”) treatment with PCZ. Shown are mean / SD values obtained from the three patients treated with PCZ. The upper limits of the healthy normal ranges are dependent on age and sex. The applicable limits according to Welsh et al. 2022 are indicated by dotted lines and the patient individual data are indicated with arrows.

[0674] Figure 9: Mean PCZ concentration and cDPP3 activity in extreme-critically ill patients with refractory shock and multi -organ failure (n = 3) after dosing with 10 mg / kg PCZ infused over two hours.

[0675] Figure 10: Schematic representation of the development and selection of anti-DPP3 antibodies using the SELMA technology. a) Myeloma cells are used for fusion with B-cells. The myeloma cells are stable transfected with a construct enabling the expression of a surface marker containing the biotin acceptor peptide sequence for site-specific biotinylation by biotin ligase (BirA). b) After fusion of these myeloma cells with the B-cells derived from an immunized mouse, the pool of hybridoma cells are biotinylated in vitro at their surface via biotin ligase. c) A separately prepared DPP3 -streptavidin conjugate is added to the pool of cells, where it binds to the biotin-residues carried by the hybridoma cells. d) During a subsequent incubation, the hybridoma cells secrete their monoclonal antibodies (mAbs). Those mAbs directed against the target protein DPP3 immediately after secretion bind to the DPP3- streptavidin conjugate nearby, whereas other mAbs from other hybridoma cells diffuse into the culture medium. e) A fluorescence-labeled anti -mouse IgG is added and binds to mAbs. “Positive” hybridoma cells now carry a fluorescence label (complex formed by biotin covalently attached at the cell surface* Streptavidin conjugated DPP3*anti-DPP3 mAb* fluorescence-labeled anti-mouse IgG); “negative” hybridoma cells do not. By fluorescence-activated cell sorting (FACS), “positive” hybridoma cells can be easily isolated.

[0676] Figure 11: Schematic representation of designs for assessment of DPP3 binding and influence on DPP3 enzymatic activity for anti-DPP3 antibodies. Examples

[0677] Example 1 - Generation of antibodies

[0678] Generation of antibodies and determination DPP3 binding ability: Several murine antibodies were produced and screened by their ability of binding human DPP3 in a specific binding assay (see Table 2).

[0679] Peptides / conjugates for immunization:

[0680] DPP3 peptides for immunization were synthesized, see Table 2, (JPT Technologies, Berlin, Germany) with an additional N-terminal cystein (if no cystein is present within the selected DPP3 -sequence) residue for conjugation of the peptides to Bovine Serum Albumin (BSA). The peptides were covalently linked to BSA by using Sulfolink-coupling gel (Perbio-science, Bonn, Germany). The coupling procedure was performed according to the manual of Perbio. Recombinant GST-hDPP3 was produced by USBio (United States Biological, Salem, MA, USA).

[0681] Immunization of mice, immune cell fusion and screening:

[0682] Balb / c mice were intraperitoneally (i.p.) injected with 84 pg GST-hDPP3 or 100 pg DPP3-peptide-BSA -conjugates at day 0 (emulsified in TiterMax Gold Adjuvant), 84 pg or 100 pg at day 14 (emulsified in complete Freund’s adjuvant) and 42 pg or 50 pg at day 21 and 28 (in incomplete Freund’s adjuvant). At day 49 the animal received an intravenous (i.v.) injection of 42 pg GST-hDPP3 or 50 pg DPP3-peptide-BSA-conjugates dissolved in saline. Three days later the mice were sacrificed, and the immune cell fusion was performed.

[0683] Splenocytes from the immunized mice and cells of the myeloma cell line SP2 / 0 were fused with 1 ml 50% polyethylene glycol for 30 s at 37°C. After washing, the cells were seeded in 96-well cell culture plates. Hybrid clones were selected by growing in HAT medium [RPMI 1640 culture medium supplemented with 20% fetal calf serum and HAT-Supplement] .

[0684] After one week, the HAT medium was replaced with HT Medium for three passages followed by returning to the normal cell culture medium.

[0685] The cell culture supernatants were primarily screened for recombinant DPP3 binding IgG antibodies two weeks after fusion. Therefore, recombinant GST-tagged hDPP3 (USBiologicals, Salem, USA) was immobilized in 96-well plates (100 ng / well) and incubated with 50 pl cell culture supernatant per well for 2 hours at room temperature. After washing of the plate, 50 pl / well peroxidase (POD)-rabbit anti mouse IgG was added and incubated for 1 h at RT. After a next washing step, 50 pl of a chromogen solution (3,7 mM o-phenylen-diamine in citrate / hydrogen phosphate buffer, 0.012% H2O2) were added to each well, incubated for 15 minutes at RT and the chromogenic reaction stopped by the addition of 50 pl 4N sulfuric acid. Absorption was detected at 490 mm.

[0686] The positive tested microcultures were transferred into 24-well plates for propagation. After retesting the selected cultures were cloned and re-cloned using the limiting-dilution technique and the isotypes were determined.

[0687] Mouse monoclonal antibody production

[0688] Antibodies raised against GST-tagged human DPP3 or DPP3 -peptides were produced via standard antibody production methods (Marx et al. 1997) and purified via Protein A. The antibody purities were > 90% based on SDS gel electrophoresis analysis.

[0689] Characterization of antibodies - binding to hDPP3 and / or immunization peptide

[0690] To analyse the capability of DPP3 / immunization peptide binding by the different antibodies and antibody clones a binding assay was performed: a) Solid phase

[0691] Recombinant GST-tagged hDPP3 (SEQ ID NO. 1) or a DPP3 peptide (immunization peptide, SEQ ID NO: 2) was immobilized onto a high binding microtiter plate surface (96-Well polystyrene microplates, Greiner Bio-One international AG, Austria, 1 pg / well in coupling buffer [50 mM Tris, 100 mM NaCl, pH7,8], Ih at RT). After blocking with 5% bovine serum albumin, the microplates were vacuum dried. h) Labelling procedure (Tracer)

[0692] 100 pg (100 pl) of the different anti-DPP3 antibodies (detection antibody, 1 mg / ml in PBS, pH 7.4) were mixed with 10 pl acridinium NHS-ester (1 mg / ml in acetonitrile, In Vent GmbH, Germany; EP 0 353 971) and incubated for 30 min at room temperature. Labelled anti-DPP3 antibody was purified by gel-filtration HPLC on Shodex Protein 5 pm KW-803 (Showa Denko, Japan). The purified labeled antibody was diluted in assay buffer (50 mmol / 1 potassium phosphate, 100 mmol / 1 NaCl, 10 mmol / 1 Na2-EDTA, 5 g / 1 bovine serum albumin, 1 g / 1 murine IgG, 1 g / 1 bovine IgG, 50 pmol / 1 amastatin, 100 pmol / 1 leupeptin, pH 7.4). The final concentration was approx. 5-7* 106relative light units (RLU) of labelled compound (approx. 20 ng labeled antibody) per 200 pl. acridinium ester chemiluminescence was measured by using a Centro LB 960 luminometer (Berthold Technologies GmbH & Co. KG). c) hDPP3 binding assay The plates were filled with 200 pl of labelled and diluted detection antibody (tracer) and incubated for 2-4 h at 2-8 °C. Unbound tracer was removed by washing 4 times with 350 pl washing solution (20 mM PBS, pH 7.4, 0.1 % Triton X-100). Well-bound chemiluminescence was measured by using the Centro LB 960 luminometer (Berthold Technologies GmbH & Co. KG).

[0693] Characterization of antibodies - hDPP3 -inhibition analysis

[0694] To analyze the capability of DPP3 inhibition by the different antibodies and antibody clones a DPP3 activity assay with known procedure (Jones et al., 1982) was performed. Recombinant GST-tagged hDPP3 was diluted in assay buffer (25 ng / ml GST-DPP3 in 50 mM Tris-HCl, pH7,5 and 100 pM ZnCE) and 200 pl of this solution incubated with 10 pg of the respective antibody at room temperature. After 1 hour of pre-incubation, fluorogenic substrate Arg-Arg-PNA (20 pl, 2mM) was added to the solution and the generation of free PNA over time was monitored using the Twinkle LB 970 microplate fluorometer (Berthold Technologies GmbH & Co. KG) at 37 °C. Fluorescence of PNA is detected by exciting at 340 nm and measuring emission at 410 nm. Slopes (in RFU / min) of increasing fluorescence of the different samples are calculated. The slope of GST-hDPP3 with buffer control is appointed as 100 % activity. The inhibitory ability of a possible capture-binder is defined as the decrease of GST-hDPP3 activity by incubation with said capture-binder in percent.

[0695] The following table represents a selection of obtained mouse monoclonal antibodies and their binding rate in Relative Light Units (RLU) as well as their relative inhibitory ability (%; Table 2). The mouse monoclonal antibodies raised against the below depicted DPP3 regions, were selected by their ability to bind recombinant DPP3 and / or immunization peptide, as well as by their inhibitory potential.

[0696] All mouse monoclonal antibodies raised against the GST-tagged, full-length form of recombinant hDPP3 show a strong binding to immobilized GST-tagged hDPP3. Mouse monoclonal antibodies raised against the SEQ ID NO: 2 peptide bind to GST-hDPP3 as well. The mouse monoclonal antibodies raised against SEQ ID NO: 2 peptide also strongly bind to the immunization peptide.

[0697] Table 2: List of mouse monoclonal antibodies raised against full-length or sequences of hDPP3 and their ability to bind hDPP3 (SEQ ID NO: 1) or immunization peptide (SEQ ID NO: 2) in RLU, as well as the maximum inhibition of recombinant GST-hDPP3.

[0698] Example 2 - Development of the humanized anti-DPP3 antibody (PCZ)

[0699] Mouse monoclonal antibodies raised against SEQ ID No.: 2 were characterized in more detail (epitope mapping, binding affinities, specificity, inhibitory potential). Here the results for murine clone 1967 of SEQ ID No.: 2 (murine anti-DPP3 antibody) are shown as an example.

[0700] Determination of murine anti-DPP3 antibody epitope on DPP3 :

[0701] For epitope mapping of murine anti-DPP3 antibody a number of N- or C-terminally biotinylated peptides were synthesized (peptides & elephants GmbH, Hennigsdorf, Germany). These peptides include the sequence of the full immunization peptide (SEQ ID No. 2) or fragments thereof, with stepwise removal of one amino acid from either C- or N-terminus (see Table 4 for a complete list of peptides).

[0702] High binding 96 well plates were coated with 2 pg Avidin per well (Greiner Bio-One international AG, Austria) in coupling buffer (500 mM Tris-HCl, pH 7.8, 100 mM NaCl). Afterwards plates were washed and filled with specific solutions of biotinylated peptides (10 ng / well; buffer - IxPBS with 0.5% BSA).

[0703] Murine anti-DPP3 antibody was labelled with a chemiluminescence label according to Example 1.

[0704] The plates were filled with 200 pl of labelled and diluted detection antibody (tracer) and incubated for 4 h at room temperature. Unbound tracer was removed by washing 4 times with 350 pl washing solution (20 mM PBS, pH 7.4, 0.1 % Triton X-100). Well-bound chemiluminescence was measured by using the Centro LB 960 luminometer (Berthold Technologies GmbH & Co. KG). Binding of murine anti-DPP3 antibody to the respective peptides is determined by evaluation of the relative light units (RLU). Any peptide that shows a significantly higher RLU signal than the unspecific binding of murine anti-DPP3 antibodyis defined as murine anti-DPP3 antibody binder. The combinatorial analysis of binding and non-binding peptides reveals the specific DPP3 epitope of murine anti-DPP3 antibody.

[0705] Determination of binding affinities using Octet:

[0706] The experiment was performed using Octet Red96 (ForteBio). Murine anti-DPP3 antibody was captured on kinetic grade anti -human Fc (AHC) biosensors. The loaded biosensors were then dipped into a dilution series of recombinant GST-tagged human DPP3 (100, 33.3, 11.1, 3.7 nM). Association was observed for 120 seconds followed by 180 seconds of dissociation. The buffers used for the experiment are depicted in Table 3. Kinetic analysis was performed using a 1: 1 binding model and global fitting.

[0707] Table 3: Buffers used for Octet measurements

[0708] Western Blot analysis of Binding specificity of murine anti-DPP3 antibody:

[0709] Blood cells from human EDTA-blood were washed (3x in PBS), diluted in PBS and lysed by repeated freeze-thaw-cycles. The blood cell lysate had a total protein concentration of 250 pg / ml, and a DPP3 concentration of 10 pg / ml. Dilutions of blood cell lysate (1:40, 1:80, 1: 160 and 1:320) and of purified recombinant human His-DPP3 (31.25-500 ng / ml) were subjected to SDS-PAGE and Western Blot. The blots were incubated in 1.) blocking buffer (IxPBS-T with 5% skim milk powder), 2.) primary antibody solution (murine anti-DPP3 antibody 1:2.000 in blocking buffer) and 3.) HRP labelled secondary antibody (goat anti mouse IgG, 1: 1.000 in blocking buffer). Bound secondary antibody was detected using the Amersham ECL Western Blotting Detection Reagent and the Amersham Imager 600 UV (both from GE Healthcare).

[0710] DPP3 inhibition assay:

[0711] To analyse the capability of DPP3 inhibition by murine anti-DPP3 antibody a DPP3 activity assay with known procedure (Jones et al., 1982) was performed as described in Example 1. The inhibitory ability murine anti-DPP3 antibody is defined as the decrease of GST-hDPP3 activity by incubation with said antibody in percent. The resulting lowered DPP3 activities are shown in an inhibition curve in Figure 1. Epitope mapping:

[0712] The analysis of peptides that murine anti-DPP3 antibody binds to and does not bind to, revealed the DPP3 sequence INPETG (SEQ ID NO: 3) as a necessary epitope for murine anti-DPP3 antibody binding (see Table 4). Similar results were gained with PCZ.

[0713] Binding affinity:

[0714] Murine anti-DPP3 antibody binds with an affinity of 2.2* 10'9M to recombinant GST-hDPP3 (kinetic curves see Figure 2).

[0715] Table 4: Peptides (SEQ ID NO. 17 to SEQ ID NO: 33) used for Epitope mapping of murine anti¬

[0716] DPP3 antibody

[0717] Specificity and inhibitory potential: The only protein detected with murine anti-DPP3 antibody as primary antibody in lysate of blood cells was DPP3 at 80 kDa (Figure 3). The total protein concentration of the lysate was 250 pg / ml whereas the estimated DPP3 concentration is about 10 pg / ml. Even though there is 25 times more unspecific protein in the lysate, murine anti-DPP3 antibody binds and detects specifically DPP3 and no other unspecific binding takes place.

[0718] Murine anti-DPP3 antibody inhibits 15 ng / ml DPP3 in a specific DPP3 activity assay with an IC50 of about 15 ng / ml (Figure 1).

[0719] Chimerization / Humanization:

[0720] The murine monoclonal antibody anti-DPP3 antibody, with the ability of inhibiting DPP3 activity by 70 %, was chosen as possible therapeutic antibody and was also used as template for chimerization and humanization.

[0721] Humanization of murine antibodies may be conducted according to the following procedure:

[0722] For humanization of an antibody of murine origin the antibody sequence is analysed for the structural interaction of framework regions (FR) with the complementary determining regions (CDR) and the antigen. Based on structural modelling an appropriate FR of human origin is selected and the murine CDR sequences are transplanted into the human FR. Variations in the amino acid sequence of the CDRs or FRs may be introduced to regain structural interactions, which were abolished by the species switch for the FR sequences. This recovery of structural interactions may be achieved by random approach using phage display libraries or via directed approach guided by molecular modelling (Almagro and Fransson, 2008. Humanization of antibodies. Front Biosci. 13:1619-33}. (Almagro 2008) The use of molecular modeling to achieve humanized antibodies may involve additional conservative amino acid substitutions at the junction of the CDR-FR. Exemplary conservative substitutions are those such as gly, ala; val, ile, leu; asp, glu; asn, gin; ser, thr; lys, arg; and phe, tyr. Additionally, back mutations may be implemented, where selection of residues outside of the region defining the specificity and thus target for back mutation (to mimic the source sequence, i.e., murine) to restore or improve the affinity of the humanized antibody. (Almagro 2008). When available, the three-dimensional models of the antibodies may be used to guide the humanization process.

[0723] With the above context, the variable region can be connected to any subclass of constant regions (IgG, IgM, IgE. IgA), or only scaffolds, Fab fragments, Fv, Fab and F(ab)2. The variable region of murine anti-DPP3 antibody in the heavy chain is shown in SEQ ID NO: 5 and the variable region of murine anti-DPP3 antibody in the light chain is shown in SEQ ID NO: 6. For chimerization and humanization a human IgG IK backbone was used. For epitope binding the Complementarity Determining Regions (CDRs) are of importance. The CDRs for the heavy chain and the light chain of the murine anti-DPP3 antibody and PCZ are shown in SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9 for the heavy chain and SEQ ID NO: 10, sequence KVS, and SEQ ID NO: 11 for the light chain, respectively.

[0724] Sequencing of the PCZ revealed an antibody heavy chain (H chain) according to SEQ ID NO: 12 and an antibody light chain (L chain) according to SEQ ID NO: 13.

[0725] Example 3 - Measurement of DPP3 concentration / activity

[0726] 3.1 Measurement of DPP3 concentration

[0727] DPP3 concentration was measured with an immunoassay using the point of care (POC) platform IB 10 from Nexus (San Diego, USA) as described in detail below (IB 10 sphingotest® DPP3). As reference the chemiluminescence sandwich-immunoassay for the quantification of DPP3 protein concentrations (DPP3-LIA, sphingotest® DPP3) (see Rehfeld et al. 2019. JALM 3(6): 943-953), was used.

[0728] In the following, details of the manufacturing and conduct of the IB 10 sphingotest® DPP3 are described:

[0729] DPP3 stock preparation

[0730] Fresh human K2-EDTA whole blood is centrifuged at 2,000 g for 5 minutes, and plasma is removed. Blood cells are washed by re-suspending in isotonic saline solution. Blood cells are lysed by repeated freeze-thaw cycles, and cell lysate is centrifuged at 20,000 g for 1 hour. The supernatant is collected and used as the DPP3 stock solution to prepare calibrators and test samples.

[0731] Calibration

[0732] The IB 10 DPP3 assay is calibrated with six levels of calibrators, which are standardized to correlate to the reference assay sphingotest® DPP3 (Rehfeld et al. 2019). The calibrators are K2-EDTA plasmabased samples with or without the DPP3 stock solution spiked. A lot-specific calibration curve is generated using a 5 parameter logistic model (MasterPlex ® ReaderFit, Hitachi).

[0733] Antibodies

[0734] Two monoclonal murine antibodies AK 2555 and AK 2553 directed against different epitopes and being able to form a sandwich with human DPP3 are produced by standard procedures from hybridoma cell lines and purified via protein A chromatography to obtain >95% purity as judged by capillary gel electrophoresis (as described in Example 1). The same antibody pair is utilized for the reference DPP3 immunoassay (sphingotest® DPP3) and the IB 10 sphingotest® DPP3 immunoassay. One monoclonal anti-DPP3 antibody AK 2553 is labelled by incubation with gold nanoparticles. The labelled antibody is washed and separated from free antibody by centrifugation. The conjugate pellet is re-suspended in storage buffer (lOmM sodium phosphate, 2% BSA, pH 7.4). The other monoclonal anti-DPP3 antibody AK 2555 is incubated with biotinamidohexanoic acid N-hydroxy succinimide ester (Sigma) to be biotinylated. After the reaction is complete, the reaction solution is dialyzed in phosphate buffered saline to retrieve the biotin-conjugated antibody without unbound biotin.

[0735] Solid phase

[0736] Streptavidin (Agilent technologies) is immobilized onto a nitrocellulose membrane (Sartorius) using a reagent dispensing system (Biodot); the streptavidin captures immunocomplexes, which are formed with the antibody conjugates and DPP3 antigen in a patient sample during assay reaction.

[0737] Assay principle

[0738] The Nexus IB10 immunochemistry system has been described previously (Song et al. 2014. Ann Clin Lab Sci. 44 (3): 254-61). The Nexus IB10 system combines chemistry with microfluidics and centrifugal flow to rapidly prepare a cell-free plasma from whole blood that can then be moved through a channel to rehydrate, solubilize, and mix with freeze-dried immunoconjugates. Through a combination of active flow and capillary action, formed complexes of the analyte, gold-conjugated antibody, and biotinylated antibody are captured at the streptavidin line. The analyte is quantitatively measured after 20 minutes with an optical signal level proportional to the analyte(s) concentration using stored lot-specific calibration parameters.

[0739] Sample testing is performed by adding 500pl (whole EDTA-blood or EDTA-plasma) to the channel inlet of the disc. The test disc includes a positive internal control to ensure that the test procedure has been conducted properly. After sample addition, the entire test is performed within the Nexus IB 10 Analyzer, which maintains a constant internal operating temperature and controls all assay operations from the whole blood separation to measurement output. 22 minutes after starting the sample run, the Nexus IB 10 Analyzer will display the results on the screen.

[0740] Assay performance

[0741] The range of DPP3 concentrations reported by the Nexus IB 10 is 5 ng / mL to 150 ng / mL. Results below or above this range will be shown as “<5 ng / mL” or “>150 ng / mL”, respectively. The LoD (Limit of Detection) of the IB 10 sphingotest® DPP3 is 5 ng / mL. LoQ (Limit of Quantitation) is the lowest DPP3 concentration that can be reproducibly measured with a total coefficient of variation of at most 20%. It was determined to be 5 ng / mL.

[0742] A comparison study was performed using matching EDTA whole blood and EDTA plasma samples. When performing a Bland-Altman analysis comparing the whole blood concentrations versus the corresponding plasma concentrations from the same subject samples (N=58), a mean difference of 9.6% (95% C.E = [6.3%- 13.0%]), and a correlation coefficient of r = 0.98 were obtained.

[0743] 3.2 DPP3 activity measurement in human plasma samples using a soluble activity assay

[0744] Enzymatic activity of DPP3 in human EDTA plasma samples was determined using a soluble activity assay with detection of a fluorescent cleavage product P-naphthylamine resulting from cleavage of a Anorogenic substrate, Arg-Arg-P-naphthylamide.

[0745] EDTA plasma samples were prediluted in neat matrix (heat inactivated human EDTA plasma pool) at equal parts (1+1). A substrate solution of the following composition was prepared: 200 pM Arg-Arg- P-naphthylamide (Bachem #K-1085), 1 mM C0CI2 (Sigma Aldrich #255599), 250 pM Amastatin (Quartett # PPI1010), 46.3 mM Tris / HCl, pH 7.8, 0.12 % Triton X-100. 5 pL of prediluted plasma samples were added to the wells of a nonbinding, uncoated, black 96 well microplate (Greiner Bio-One #7635165) and 95 pL of substrate solution were added leading to a final sample dilution of 1:40 relative to the neat sample. The plate was immediately transferred to incubate in a multi-mode microplate reader (SpectraMax iD5) preheated to 37°C. Kinetic measurement of the DPP3 enzymatic reaction, e.g. the cleavage of Arg-Arg-P-naphthylamide, was done with an excitation wavelength of 340 nm and emission at 410 nm in intervals of 2 minutes for a total duration of maximal 40 minutes. Velocity of the DPP3 reaction was determined on the individual sample’s linear phase, which started approximately 20 minutes after start of the reaction and lasted till the end of measurement (maximal 40 minutes after start of the reaction) (curve fitting with R2 > 0.98) and red out as relative ftuorescence units (RFU) per minute. The measured RFU were converted into the molar amount of P-naphthylamine as follows: First, serial dilutions of P-naphthylamine were prepared, and their RFU were measured within the same plate as samples. By linear regression of these data (forced through origin), it was determined how much RFU are generated per amount of P-naphthylamine, e.g. RFU / pmol. Using this factor, the activity measured for the samples in RFU / min was converted into pmol / min.

[0746] All samples as well as dilutions of P-naphthylamine were measured in duplicate.

[0747] Blood based samples contain compounds that unspecifically interfere to a certain extent with the DPP3 activity assay and lead to an underestimation of the actual DPP3 activity. To correct for such unspecific interference a spike and recovery test based on human, native DPP3 was performed per sample in the activity assay. To this end, 50 pL of a 100 ng / mL solution of purified, human, native DPP3 (“spike solution”) were added to 50 pL of each sample, 5 pL of this mix were pipetted onto the assay plate and 95 pL substrate solution were added, as described above, and the enzymatic DPP3 activity was measured, as described above. Separately, the DPP3 activity of the “spike solution” was determined from a “spike in heat-inactivated EDTA plasma” sample, where 50 pL spike solution were diluted with 50 pL heat-inactivated EDTA plasma. From this dilution 5 pL were mixed with 95 pL substrate solution, and DPP3 activity was measured, as described above.

[0748] In the absence of a specific inhibitor of DPP3, such as for instance PCZ, the calculation of unspecific interference from the spike and recovery test would be straightforward by simply comparing the DPP3 activity of the spike solution with the difference of the DPP3 activities measured in the spiked and unspiked sample. However, in the presence of a specific inhibitor of DPP3 it is not, since the inhibitory effect of this inhibitor adds to the unspecific interference. Thus, a modified calculation of the recovery was established: A partial non-competitive inhibitor model (Masson and Mukhametgalieva 2023) was applied to calculate the “unspecifically interfered” DPP3 concentrations in unspiked and spiked sample. The model allows to calculate the “unspecifically interfered” DPP3 concentrations via activity (velocity) measured in absence or presence of a specific inhibitor of DPP3 with predefined protocol specific parameters (see further below). Calculations were done using Excel Solver add-on (GRG-non-linear algorithm).

[0749] Equation 1: (Masson and Mukhametgalieva 2023) For calculation of the enzyme concentration [E] .

[0750] In the absence of a specific inhibitor of DPP3, the inhibitor concentration in the equation | Z| is set to 0, and only the following physical values are entered in the equation: v: Velocity of enzyme (here DPP3) reaction, represents activity [pmol / min]

[0751] Kcat'. Turnover rate of the enzyme (here DPP3) / catalytic constant, previously determined = 0.8515 [pmol min1pM1] Ks: Enzyme (here DPP3) substrate (here Arg-Arg-P-naphthylamide) dissociation constant (Km: Michaelis-Menten constant), previously determined = 0.01874 mM

[0752] [E] Enzyme concentration, (here the “unspecifically interfered” DPP3) (final concentration in the activity assay) [pM]

[0753] [S] Substrate concentration (here Arg-Arg-P-naphthylamide), here 200 pM

[0754] The constant parameters were determined by solving the equation 1 for each constant with different level of E or S as follows:

[0755] The parameters Kcat, Ki, B and Ks were estimated by measuring the activity of DPP3 (100 ng / mL spiked in heat inactivated human EDTA-plasma pool). The measurements were done with 8 different substrate concentrations (0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5 and 0.6 mM (final concentration in the assay)) to ensure correct estimation of Ks.

[0756] For both, the unspiked and the spiked samples, the “unspecifically interfered” DPP3 concentration was estimated using equation 1 by varying the DPP3 concentration (using the “Solver” function in MS Excel) to approximate the most accurate solution (in the absence of PCZ, [I] was set to 0).

[0757] It is assumed that the estimated DPP3 concentrations in the unspiked and the spiked sample are similarly affected by unspecific interference.

[0758] With the estimated “unspecifically interfered” DPP3 concentrations of the unspiked and the spiked sample derived from equation 1 and the known DPP 3 concentration of the spike solution, the recovery could now be calculated. Recovery was calculated by subtracting the estimated “unspecifically interfered” DPP3 concentration from the unspiked sample from the estimated “unspecifically interfered” DPP3 concentration of its corresponding spiked sample; the difference is then divided by the DPP3 concentration of the spike solution and multiplied by 100%.

[0759] Any reduction in recovery is indicative of unspecific interference within the sample, and this quantified recovery is subsequently used to correct the measured DPP3 activity of the unspiked sample. The correction was performed by dividing the measured DPP3 activity of the unspiked sample by the corresponding recovery of the spiked DPP3 concentration and then multiplying by 100%.

[0760] The corrected activities [pmol min1] now represented the enzyme activity of the diluted sample in the assay solution after accounting for unspecific interference. To determine the enzyme activity in the undiluted sample, the corrected activity was multiplied by the total dilution factor of 40. Finally, by multiplying the resulting activity in the sample by a factor of 10,000, the enzyme activity in 1 liter of undiluted sample was calculated, expressed as pmol min1L1, which is equivalent to [U L1].

[0761] Example 4 - Administration of humanized anti-DPP3 antibody (procizumab, PCZ) in extreme- critically ill patients with refractory shock and multi-organ failure

[0762] PCZ was administered in three extreme-critically ill patients with refractory shock and multi-organ failure.

[0763] Patient description'.

[0764] The patients included in this study constitute a specific patient population, for which treatment with PCZ and resulting outcome have not been investigated in the art so far. This population can be termed “extreme-critically ill patients”. This term is defined by the following features: Extreme-critically ill patient according to the present invention is defined as a patient who has a 24-hour probability of mortality of at least 70% and / or has organ dysfunction requiring mechanical or pharmacological support of at least two organs.

[0765] “Critically ill” means that said patient is suffering from an acute disease or acute condition which is life- threatening and in which death is possible or imminent. In a specific embodiment said critically ill patient is an ICU patient.

[0766] The patient has a 24-hour probability of mortality of at least 70%, preferably of at least 80%, more preferably of at least 90%, most preferably of at least 95% and / or characterized by organ dysfunction requiring mechanical or pharmacological support of at least two organs . Two women (patients PCZ 01 , 64 years old, and PCZ 03, 80 years old) and one man (patient PCZ_02, 84 years old) were treated. All three patients had pre-existing conditions and were in utmost critical condition with signs of multi -organ failure, with patient PCZ_01 and PCZ_02 suffering from combined septic and cardiogenic shock (AMI in patient PCZ 01 was untreated due to hemodynamic instability) and patient PCZ 03 from right-sided cardiogenic shock. All three were suffering from renal failure, in need of renal replacement therapy, two out of three (patients PCZ_01 and PCZ_02) were intubated and under invasive mechanical ventilation. All three patients were assessed and approved for treatment with PCZ by an independent international patient selection board, which considered patient PCZ 01 as having a 24-hour probability of mortality of >90%, and patients PCZ_02 and PCZ_03 as already having entered the process of dying.

[0767] Intervention'. Patients received a single dose of PCZ infusion at a dose of 10 mg / kg body weight over a 2-hour period.

[0768] PCZ was administered using a dedicated lumen of a central venous catheter, separately from any concomitant drugs, between 1 and 3 days after transfer to intensive care unit (ICU) .

[0769] Follow -up

[0770] The patients were observed for a period of 6 days after administration of PCZ, and several blood samples were collected in this period for the assessment of PCZ concentration and cDPP3 activity.

[0771] Results'.

[0772] The therapy was well tolerated in all three patients, no immediate adverse reactions were noted, and no adverse events were associated with PCZ.

[0773] Clinical and laboratory data recorded for the patients immediately prior to the administration of PCZ as well as after 24 and 48 hours thereafter are summarized in Table 5.

[0774] Prior to administration of PCZ all patients suffered from multi -organ failure: Affected organs included the cardiovascular system (mean norepinephrine dosing 0.9 pg / kg / min), the lung (mean Horovitz-Index (P / F ratio) 118 mmHg), as well as the kidney (mean creatinine concentration 2.3 mg / dL) and required multi -organ support.

[0775] Table 5: Clinical and laboratory patient characteristics prior to and after treatment with PCZ.

[0776] Surprisingly, despite the patients being extreme-critically ill, treatment with PCZ resulted in a rapid improvement with respect to several relevant clinical features.

[0777] Administration of PCZ led to a successful treatment of shock, e.g., a significant reduction of required dosing of the vasopressor norepinephrine. Prior to the treatment with PCZ, patient PCZ 01, patient PCZ_02, and patient PCZ_03 received norepinephrine doses of 0.8, 1.4, and 0.6 pg / kg / min (mean: 0.9 pg / kg / min), respectively. One day after the PCZ treatment, the required doses of norepinephrine dropped to 67%, and after 48 hours down to 31%, compared to the starting dose.

[0778] Lactate is an established biomarker in critical care. Blood lactate levels are increased due to inadequate tissue oxygenation and other processes (Bakker, Nijsten, and Jansen 2013). Increased lactate levels reflect increased morbidity and high mortality risk, and decrease of lactate levels over time has been shown to be associated with a favorable outcome in diverse critically ill populations including post cardiac surgery, trauma patients, sepsis and septic shock, cardiogenic shock, cardiac arrest, respiratory failure, and others (Vincent et al. 2016).

[0779] In the present study, all three patients presented with highly elevated levels of lactate, e.g., above the established clinical cut-off of 2 mmol / L (Vieira, Petrova, and Moura 2022). Upon treatment with PCZ, levels steadily declined and reached non-critical concentrations within 48 hours after start of treatment. Figure 5. Median levels of lactate in healthy subjects have been reported to be 1.03 mmol / L (Sung et al. 2016).

[0780] Interleukin-6 (IL-6) is a biomarker of systemic inflammation which has been shown in a diverse population of critically ill patients to reflect the extent of organ dysfunction, need for organ-support therapies, hemodynamic dysfunction characterized by reduced systolic and diastolic arterial pressure, coagulopathy, and hepatic, and renal dysfunction (Picod et al. 2022).

[0781] The extreme-critically patients of the present study exhibited significantly elevated IL-6 levels prior to treatment with PCZ. These levels steadily declined over the observation period of 48 hours (Figure 6). For the general adult population, a median of 2.1 pg / mL and an upper reference range of 5.9 pg / mL have been reported earlier (Alende-Castro, V, Alonso-Sampedro, M, Gude, F, Gonzalez-Quintela 2021).

[0782] The effect of PCZ on the kidney function was assessed. All patients suffered from severely impaired kidney function, likely due to acute kidney injury, as reflected by high levels of blood creatinine and a very low glomerular filtration rate (GFR). They all required renal replacement therapy (RRT). Upon treatment with PCZ, both measures of kidney function improved steadily (Figure 7 A and Figure 7 B). Kidney function is considered impaired when the GFR is below 90 mL / min / 1.73 m2(Levey et al. 2005). Reference intervals for serum creatinine concentrations have been reported as 0.72-1.18 mg / dL (2.5thto 97thpercentile) for adult men and 0.55-1.02 mg / dL (2.5thto 97thpercentile) for adult women (Ceriotti et al. 2008).

[0783] Additionally, the effect of PCZ on the heart function was assessed by measuring N-terminal prohormone of brain natriuretic peptide (NT-proBNP). NT-proBNP is an inactive prohormone fragment stemming from the same precursor peptide as the active B-type natriuretic peptide (BNP). Both BNP and NT- proBNP levels in the blood are used for screening, diagnosis of acute congestive heart failure (CHF) and may be useful to establish prognosis in heart failure, as both markers are typically higher in patients with worse outcome (Samad, Malempati, and Restini 2023). The plasma concentrations of both BNP and NT- proBNP are also typically increased in patients with asymptomatic or symptomatic left ventricular dysfunction and are associated with coronary artery disease, myocardial ischemia, and severity of aortic valve stenosis (Samad, Malempati, and Restini 2023). All three patients presented with supranormal levels of NT-proBNP, indicating an impaired heart function. Upon treatment with PCZ, levels of NT- proBNP declined steadily (Figure 8). Normal reference ranges of NT-proBNP are dependent on age and sex and have been reported in detail by (Welsh et al. 2022), see Table 6:

[0784] Table 6: Reference values for the 99thpercentile of NT-proBNP among 18,356 participants with no cardiovascular disease. [07...

Claims

1. T75380WO BOEHMERT & BOEHMERTClaims1. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock, wherein said patient is an extreme-critically ill patient.

2. Anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function, wherein said shock patient has acute kidney injury (AKI).

3. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock according to claim 1 or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to claim 2, wherein said shock is selected from the group comprising hypovolemic shock, cardiogenic shock, obstructive shock and distributive shock, or wherein said shock patient has combined cardiogenic and septic shock.

4. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to any of claims 1 to 3, wherein said patient has a 24-hour probability of mortality of at least 70%, preferably of at least 80%, more preferably of at least 90%, most preferably of at least 95% and / or has organ dysfunction requiring mechanical or pharmacological support of at least two organs.

5. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to claim 4, wherein said organ dysfunction is defined as cardiovascular dysfunction, respiratory dysfunction (lung), kidney dysfunction, liver dysfunction, central nervous system dysfunction or dysfunction of the coagulation system.

6. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for usein therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to claim 4 or 5, wherein said organ support is selected from kidney support, in particular renal replacement therapy, respiratory (lung) support, cardiovascular support and / or liver support.

7. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to any of claims 1 to 6, wherein the level of DPP3 in a sample of a bodily fluid of said patient is above a threshold and wherein said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample.

8. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to claim 7, wherein said threshold of DPP3 concentration is between 20 and 120 ng / mL, more preferred between 25 and 100 ng / mL, even more preferred between 30 and 80 ng / mL, even more preferred between 35 and 60 ng / mL, even more preferred between 40 and 50 ng / ml most preferred said threshold is 40 ng / mL.

9. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to claim 7, wherein said threshold of enzymatic activity of DPP3 is between 1.76 and 9.68 pmol / (min*L), more preferred between 2.15 and 9.09 pmol / (min*L), even more preferred between 2.55 and 6.51 pmol / (min*L), even more preferred between 2.94 and 4.93 pmol / (min*L), even more preferred between 3.34 and 4.13 pmol / (min*L), most preferred said threshold is 3.34 pmol / (min*L).

10. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to any of claims 7 to 9, wherein said bodily fluid is selected from the group comprising whole blood, serum and plasma.

11. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to any of claims 1 to 10, wherein the patient is characterized by a level of lactate of 2 mmol / 1 or higher and / or a level of IL-6 of 5.9 pg / ml or higher and / or a level of CRP of 1 mg / dL or higher and / or a level of PCT of 0.5 ng / ml or higher and / or a level of bio- ADM of 70 pg / ml or higher and / or serum creatinine of 1.18 mg / dL (for men) and 1.02 mg / dL (for women) and / or a supranormal level of NT-proBNP in a sample of bodily fluid of said patient.

12. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to any of claims 1 to 11, wherein the heavy chain comprises at least one CDR, preferably at least three CDRs selected from the group comprising:SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / orSEQ ID No. 9: ARNYSYDY. and wherein the light chain comprises at least one CDR, preferably at least three CDRs selected from the group comprising:SEQ ID No. 10: RSLVHSIGSTY,CDR2: KVS and / orSEQ ID No. 11 : SQSTHVPWT or alternatively a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

13. Anti-DPP3 antibody thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to any of claims 1 to 12, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody comprises the following sequences:SEQ ID No. 12MDPKGSLSWRILLFLSLAFELSYGQITLKESGPTLVKPTQTLTLTCTFSGFSLSTSG MSVGWIRQPPGKALEWLAHIWWNDNKSYNPALKSRLTITRDTSKNQVVLTMTN MDPVDTGTYYCARNYSYDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG and / orSEQ ID No. 13METDTLLLWVLLLWVPGSTGDIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGST YLYWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVY YCSQSTHVPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLS SPVTKSFNRGEC or alternatively sequences that are > 80% identical, preferably > 90% identical, preferably > 95% identical, preferably > 98% identical, preferably > 99% identical thereto, wherein the heavy chain comprises at least one CDR selected from the group comprising SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / orSEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR selected from the group comprising:SEQ ID No. 10: RSLVHSIGSTY,CDR2: KVS and / orSEQ ID No. 11: SQSTHVPWT, or a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

14. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to any of claims 1 to 13, wherein an anti-DPP3 antibody is used and wherein said anti -DPP3 antibody is administered at a dose between 1 to 30 mg / kg bodyweight, preferably 2 to 20 mg / kg bodyweight, preferably 3 to 15 mg / kg bodyweight, preferably 5 to 12 mg / kg body weight, preferably 7 to 10 mg / kg body weight and most preferred 10 mg / kg body weight.

15. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to any of claims 1 to 14, wherein an anti -DPP3 antibody is used and wherein said anti-DPP3 is administered for 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours.

16. Anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to any of claims 1 to 15, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody is administered at a dose of 10 mg / kg bodyweight and administered as infusion over 1 to 3 hours, preferably 1.5 to 2.5 hours and most preferred 2 hours.

17. Anti-DPP3 antibody thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to any of claims 1 to 12 or 14 to 16, wherein an anti-DPP3 antibody is used and wherein said anti-DPP3 antibody comprises the following sequences:SEQ ID No. 34 QITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGMSVGWIRQPPGKALEWLAHIWW NDNI<SYNPAL1<SRLTITRDTSI<NQVVLTMTNMDPVDTGTYYCARNYSYDYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGAL TSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWE SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPG and / orSEQ ID No. 35DIVMTQTPLSLSVTPGQPASISCKSSRSLVHSIGSTYLYWYLQKPGQSPQLLIYKVS NRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPWTFGGGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC or alternatively sequences that are > 80% identical, preferably > 90% identical, preferably > 95% identical, preferably > 98% identical, preferably > 99% identical thereto, wherein the heavy chain comprises at least one CDR selected from the group comprising SEQ ID No. 7: GFSLSTSGMS, SEQ ID No. 8: IWWNDNK and / orSEQ ID No. 9: ARNYSYDY and wherein the light chain comprises at least one CDR selected from the group comprising:SEQ ID No. 10: RSLVHSIGSTY,CDR2: KVS and / orSEQ ID No. 11 : SQSTHVPWT, or a sequence wherein one or two amino acids are exchanged from the original CDR sequences in at least one of the CDR sequences provided that the binding affinity to DPP3 is diminished by not more than 20%.

18. An anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for use in therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to any of claims 1 to 17, wherein said shock is refractory shock.

19. An anti-DPP3 antibody or fragment thereof binding to DPP3 for the treatment of shock in a patient having shock or anti-DPP3 antibody or fragment thereof binding to DPP3 for usein therapy in a critically ill patient with shock for improvement or stabilization of kidney function according to any of claims 1 to 18, wherein said patient is characterized by a level of DPP3 in a sample of a bodily fluid of said patient above a threshold, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and wherein the level of DPP3 is determined by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3.

20. A method comprising determining the level of DPP3 in a sample of a bodily fluid of a patient having shock, wherein said patient is an extreme-critically ill patient, by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and, if said level of DPP3 in said sample is above a threshold, said patient is requiring administration of an anti-DPP3 antibody or fragment thereof for the treatment of said shock.

21. A method for the treatment of shock in an extreme-critically ill patient having shock, comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof, wherein said extreme-critically ill patient is characterized by a level of DPP3 in a sample of a bodily fluid of said patient above a threshold, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and wherein the level of DPP3 is determined by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3.

22. A method for the treatment of shock in an extreme-critically ill patient having shock, comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof, wherein the level of DPP3 determined in a sample of a bodily fluid of said patient by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3 is above a threshold,and wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample.

23. A method comprising determining the level of DPP3 in a sample of a bodily fluid of an extreme-critically ill patient having shock by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and, if said level of DPP3 in said sample is above a threshold, administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof.

24. A method comprising determining the level of DPP3 in a sample of a bodily fluid of a patient with shock and having acute kidney injury (AKI) by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and, if said level of DPP3 in said sample is above a threshold, categorizing the patient as being critically ill and as requiring administration of an anti-DPP3 antibody or fragment thereof for use in therapy of said patient and for improvement or stabilization of kidney function in said patient.

25. A method for the treatment of shock in a critically ill patient with shock and having acute kidney injury (AKI), comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof, wherein said critically ill patient is characterized by a level of DPP3 in a sample of a bodily fluid of said patient above a threshold, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and wherein the level of DPP3 is determined by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3.

26. A method for the treatment of shock in a critically ill patient with shock and having acute kidney injury (AKI), comprising administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof,wherein the level of DPP3 determined in a sample of a bodily fluid of said patient by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3 is above a threshold, and wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample.

27. A method comprising determining the level of DPP3 in a sample of a bodily fluid of a critically ill patient with shock and having acute kidney injury (AKI) by a method comprising contacting said sample of bodily fluid with a capture binder that binds specifically to DPP3, wherein in particular said level of DPP3 is the DPP3 concentration and / or the enzymatic activity of cDPP3 in said sample, and, if said level of DPP3 in said sample is above a threshold, administering to said subject an effective amount of an anti-DPP3 antibody or fragment thereof.

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