Abstract: Herein is reported a method for obtaining a polypeptide in monomeric form comprising the steps of a) providing a solution comprising the polypeptide in monomeric form and in aggregated form, wherein the ratio of monomeric to aggregated form is 4:1 or less as determined by size exclusion chromatography, b) performing a mixed-mode chromatography in bind-and-elute mode, or a hydrophobic interaction chromatography in flow-through mode, or a size-exclusion chromatography, and c) performing a weak cation exchange chromatography in bind-and-elute mode or flow-through mode, and thereby obtaining the polypeptide in monomeric form.

Abstract: A micro fluid filtration system (100) preferably for increasing the concentration of components contained in a fluid sample has a fluid circuitry (1). The fluid circuitry (1) comprises the following elements: A tangential flow filtration element (7) capable for separating the fluid sample into a retentate stream and a permeate stream upon passage of the fluid, an element for pumping (3) for creating and driving a fluid flow through the fluid circuitry (1) and at least one element for obtaining information about the properties of the fluid sample within the circuitry. The circuitry further comprises a plurality of conduits (24) connecting the elements of the fluid circuitry (1) through which a fluid stream of the fluid sample is conducted. The circuitry (1) has a minimal working volume of at most 5 ml, which is the minimal fluid volume retained in the elements and the conduits (24) of the circuitry (1) such that the fluid can be recirculated in the circuitry (1) without pumping air through the circuitry (1).

Abstract: Herein is reported a method for obtaining a polypeptide in monomeric form comprising the steps of a) providing a solution comprising the polypeptide in monomeric form and in aggregated form, wherein the ratio of monomeric to aggregated form is 4:1 or less as determined by size exclusion chromatography, b) performing a mixed-mode chromatography in bind-and-elute mode, or a hydrophobic interaction chromatography in flow-through mode, or a size-exclusion chromatography, and c) performing a weak cation exchange chromatography in bind-and-elute mode or flow-through mode, and thereby obtaining the polypeptide in monomeric form.

Abstract: Herein is reported a method for obtaining a polypeptide in monomeric form comprising the steps of a) providing a solution comprising the polypeptide in monomeric form and in aggregated form, wherein the ratio of monomeric to aggregated form is 4:1 or less as determined by size exclusion chromatography, b) performing a mixed-mode chromatography in bind-and-elute mode, or a hydrophobic interaction chromatography in flow-through mode, or a size-exclusion chromatography, and c) performing a weak cation exchange chromatography in bind-and-elute mode or flow-through mode, and thereby obtaining the polypeptide in monomeric form.

Abstract: A micro fluid filtration system (100) preferably for increasing the concentration of components contained in a fluid sample has a fluid circuitry (1). The fluid circuitry (1) comprises the following elements: A tangential flow filtration element (7) capable for separating the fluid sample into a retentate stream and a permeate stream upon passage of the fluid, an element for pumping (3) for creating and driving a fluid flow through the fluid circuitry (1) and at least one element for obtaining information about the properties of the fluid sample within the circuitry. The circuitry further comprises a plurality of conduits (24) connecting the elements of the fluid circuitry (1) through which a fluid stream of the fluid sample is conducted. The circuitry (1) has a minimal working volume of at most 5 ml, which is the minimal fluid volume retained in the elements and the conduits (24) of the circuitry (1) such that the fluid can be recirculated in the circuitry (1) without pumping air through the circuitry (1).

Abstract: A micro flow filtration system comprises a fluid circuitry (3) and a first reservoir (1) outside the circuitry (3) suitable for containing a fluid. The fluid circuitry (3) comprises a tangential flow filtration module (10) capable of separating the fluid sample into a retentate stream and a permeate stream upon passage of the fluid sample into the tangential flow filtration module (10) through an inlet feed (18). The fluid circuitry (3) further comprises a second reservoir (2) integrated in the fluid circuitry (3), a pump (5) for creating and driving a fluid flow, optionally at least one pressure sensor (6, 7 or 8) for acquiring and detecting data about the fluid sample, optionally a pressure regulator (9) for regulating the flow in the fluid circuitry (3) and a plurality of conduits (22) forming the fluid circuitry (3) together with the second reservoir (2), the TFF-module (10), the pump (5), the pressure sensor (6, 7 or 8) (if present) and the pressure regulator (9) (if present).

Abstract: A micro fluid filtration system (100) preferably for increasing the concentration of components contained in a fluid sample has a fluid circuitry (1). The fluid circuitry (1) comprises the following elements: A tangential flow filtration element (7) capable for separating the fluid sample into a retentate stream and a permeate stream upon passage of the fluid, an element for pumping (3) for creating and driving a fluid flow through the fluid circuitry (1) and at least one element for obtaining information about the properties of the fluid sample within the circuitry. The circuitry further comprises a plurality of conduits (24) connecting the elements of the fluid circuitry (1) through which a fluid stream of the fluid sample is conducted. The circuitry (1) has a minimal working volume of at most 5 ml, which is the minimal fluid volume retained in the elements and the conduits (24) of the circuitry (1) such that the fluid can be recirculated in the circuitry (1) without pumping air through the circuitry (1).

Abstract: Herein is reported a method for obtaining a polypeptide in monomeric form comprising the steps of a) providing a solution comprising the polypeptide in monomeric form and in aggregated form, wherein the ratio of monomeric to aggregated form is 4:1 or less as determined by size exclusion chromatography, b) performing a mixed-mode chromatography in bind-and-elute mode, or a hydrophobic interaction chromatography in flow-through mode, or a size-exclusion chromatography, and c) performing a weak cation exchange chromatography in bind-and-elute mode or flow-through mode, and thereby obtaining the polypeptide in monomeric form.

Abstract: Herein is reported a method for obtaining a polypeptide in monomeric form comprising the steps of a) providing a solution comprising the polypeptide m monomeric form and m aggregated form, wherein the ratio of monomeric to aggregated form is 4:1 or less as determined by size exclusion chromatography, b) performing a mixed-mode chromatography in bind-and-elute mode, or a hydrophobic interaction chromatography in flow-through mode, or a size-exclusion chromatography, and c) performing a weak cation exchange chromatography in bind-and-elute mode or flow-through mode, and thereby obtaining the polypeptide m monomeric form.

Abstract: A flow filtration system (1) for concentration of components contained in a fluid sample comprises a fluid channel (2) being formed by two conduits (4, 5) allowing a bidirectional flow of the fluid sample through the fluid channel (2), a tangential flow filtration module (3) and at least two pairs (8, 9) of piston pumps (6) each having two piston pumps (6), wherein the piston pumps (6) having a piston swept volume forming a reservoir being able to contain the fluid for volumes of up to 100 ml. The tangential flow filtration module (3) is located in the fluid channel (2) so that a fluid flowing through the channel (2) passes through the filtration module (3). At each end (10, 11) of the channel (2) one piston pump (6) of each pair (8, 9) of piston pumps (6) is located in such a manner that the piston pumps are fluidically connected in parallel.

Abstract: Herein is reported a method for the recombinant production of a polypeptide, which comprises the dipeptide AR, characterized in that the method comprises the recovering of the polypeptide from the cells or the cultivation medium of a cultivation of a cell comprising a nucleic acid encoding the polypeptide and thereby producing the polypeptide, whereby the dipeptide AR comprised in the polypeptide is encoded by the oligonucleotide gca cgt, or the oligonucleotide gcg cgt, or the oligonucleotide gcc cgt.

Abstract: The current invention reports a method for the purification of a not-glycosylated, heterologous polypeptide, which has been recombinantly produced in a prokaryotic cell, wherein the method comprises three chromatography steps of which the first chromatography step selected from i) hydrophobic charge induction chromatography, or ii) hydrophobic interaction chromatography, or iii) affinity chromatography, or iv) ion exchange chromatography, the second chromatography step is selected from i) anion exchange chromatography, or ii) cation exchange chromatography, or iii) hydroxylapatite chromatography, or iv) hydrophobic interaction chromatography, and the a third chromatography step is selected from i) hydrophobic charge induction chromatography, or ii) anion exchange chromatography, or iii) cation exchange chromatography, or iv) hydrophobic interaction chromatography, whereby the first chromatography step is an affinity chromatography in case of polypeptides capable of interacting with metal ligands, the second c

Abstract: A method for producing a lipid particle comprising the following: i) providing a first solution comprising denatured apolipoprotein, ii) adding the first solution to a second solution comprising at least two lipids and a detergent but no apolipoprotein, and iii) removing the detergent from the solution obtained in ii) and thereby producing a lipid particle.

Abstract: A flow filtration system (1) for concentration of components contained in a fluid sample comprises a fluid channel (2) being formed by two conduits (4, 5) allowing a bidirectional flow of the fluid sample through the fluid channel (2), a tangential flow filtration module (3) and at least two pairs (8, 9) of piston pumps (6) each having two piston pumps (6), wherein the piston pumps (6) having a piston swept volume forming a reservoir being able to contain the fluid for volumes of up to 100 ml. The tangential flow filtration module (3) is located in the fluid channel (2) so that a fluid flowing through the channel (2) passes through the filtration module (3). At each end (10, 11) of the channel (2) one piston pump (6) of each pair (8, 9) of piston pumps (6) is located in such a manner that the piston pumps are fluidically connected in parallel.

Abstract: Herein is reported a method for the recombinant production of a polypeptide, which comprises the tripeptide QKK, characterized in that the method comprises the step of recovering the polypeptide from the cells or the cultivation medium of a cultivation of a cell comprising a nucleic acid encoding the polypeptide and thereby producing the polypeptide, whereby the tripeptide QKK comprised in the polypeptide is encoded by the oligonucleotide cag aaa aaa or the oligonucleotide caa aag aaa.

Abstract: Herein is reported a method for the recombinant production of a polypeptide, which comprises the dipeptide AR, characterized in that the method comprises the recovering of the polypeptide from the cells or the cultivation medium of a cultivation of a cell comprising a nucleic acid encoding the polypeptide and thereby producing the polypeptide, whereby the dipeptide AR comprised in the polypeptide is encoded by the oligonucleotide gca cgt, or the oligonucleotide gcg cgt, or the oligonucleotide gcc cgt.

Abstract: A micro flow filtration system comprises a fluid circuitry (3) and a first reservoir (1) outside the circuitry (3) suitable for containing a fluid. The fluid circuitry (3) comprises a tangential flow filtration module (10) capable of separating the fluid sample into a retentate stream and a permeate stream upon passage of the fluid sample into the tangential flow filtration module (10) through an inlet feed (18). The fluid circuitry (3) further comprises a second reservoir (2) integrated in the fluid circuitry (3), a pump (5) for creating and driving a fluid flow, optionally at least one pressure sensor (6, 7 or 8) for acquiring and detecting data about the fluid sample, optionally a pressure regulator (9) for regulating the flow in the fluid circuitry (3) and a plurality of conduits (22) forming the fluid circuitry (3) together with the second reservoir (2), the TFF-module (10), the pump (5), the pressure sensor (6, 7 or 8) (if present) and the pressure regulator (9) (if present).

Abstract: A micro fluid filtration system (100) preferably for increasing the concentration of components contained in a fluid sample has a fluid circuitry (1). The fluid circuitry (1) comprises the following elements: A tangential flow filtration element (7) capable for separating the fluid sample into a retentate stream and a permeate stream upon passage of the fluid, an element for pumping (3) for creating and driving a fluid flow through the fluid circuitry (1) and at least one element for obtaining information about the properties of the fluid sample within the circuitry. The circuitry further comprises a plurality of conduits (24) connecting the elements of the fluid circuitry (1) through which a fluid stream of the fluid sample is conducted. The circuitry (1) has a minimal working volume of at most 5 ml, which is the minimal fluid volume retained in the elements and the conduits (24) of the circuitry (1) such that the fluid can be recirculated in the circuitry (1) without pumping air through the circuitry (1).

Abstract: Herein is reported a method for obtaining a polypeptide by an immobilized metal ion affinity chromatography from a pro-polypeptide that comprise at its N- or C-terminus an metal ion affinity chromatography tag and a protease cleavage site comprising the step of recovering the polypeptide from the immobilized metal ion affinity chromatography column by incubating the bound pro-polypeptide with a protease, whereby the immobilized metal ion affinity chromatography material has been washed at least once with an urea solution.

Abstract: A pro-polypeptide which is useful for the expression of a polypeptide of interest in a prokaryotic cell. Therefore the pro-polypeptide is fused to the N-terminus of the polypeptide of interest. The pro-polypeptide as reported herein provides for improved expression yields and improves the handling of the fusion polypeptide (downstream processing, purification). For example, efficient endotoxin removal is effected while the protein of interest comprising the pro-polypeptide is bound e.g. to an affinity chromatography material. Thereafter the pro-polypeptide can efficiently be cleaved from the polypeptide of interest by the incorporated protease cleavage site with the cognate protease.