Hoffmann-la Roche Inc.

Roche Diagnostics Operations, Inc.

Roche Diagnostics Gmbh

F. Hoffmann-la Roche Ag

Leopold Von Proff

Michael Schraeml

Uwe Kobold

Herbert Von Der Eltz

Hans-Peter Josel

Christoph Seidel

Rupert Herrmann

Erasmus Huber

Michael Gerg

Thomas Meier

Michael Grol

Klaus Hallermayer

Monika Soukupova

Thomas Duelffer

Klaus Hirzel

Caroline Dorothea Hojer

Ulrike Kurtkaya

Susanne Feldmann

Sonja Fiesel

Eramus Huber

Nicole Horn

Marion Herrmann

Angela Puhlmann

Stefanie Kern

Apostolos Kyriatsoulis

Christina Porzig

Lars Hillringhaus

The present invention relates to a monoclonal antibody capable of binding to biotin. In one embodiment the monoclonal antibody according to the invention also does not bind to a biotin moiety on a biotinylated molecule, wherein the biotin moiety is attached to the molecule via the carbon atom of the carboxyl function of the valeric acid moiety of biotin. Also disclosed is a method for generation of an antibody as discloed herein. The monoclonal antibody according to the invention is of specific use in a method for measuring an analyte in a sample, wherein a (strept)avidin/biotin pair is used to bind a biotinylated analyte specific binding agent to a (strept)avidin coated solid phase.

Biotin-specific monoclonal antibody and use thereof

The current invention reports a method for producing an antibody comprising the steps of a) providing a plurality of hybridoma cells each expressing an antibody, b) determining the time dependent amount of said antibody bound to the respective antigen by surface plasmon resonance at different temperatures and different antibody concentrations, c) calculating with the time dependent amount determined in b) based on equations (II) to (XIII) at least the thermodynamic parameters (i) standard association binding entropy (&#x394;S&#xb0;&#x2021;ass), (ii) standard dissociation binding entropy (&#x394;S&#xb0;&#x2021;diss), (iii) standard binding entropy (&#x394;S&#xb0;), (iv) free standard binding enthalpy (&#x394;G&#xb0;), (v) standard dissociation free binding enthalpy (&#x394;G&#xb0;&#x2021;diss), (vi) standard association free binding enthalpy (&#x394;G&#xb0;&#x2021;ass), (vii) &#x2212;T&#x394;S&#xb0;, (viii) dissociation rate constant k, (ix) equilibrium binding constant K, and (x) association rate constant k, d) selecting a hybridoma cell producing an antibody with at least two of the following: i) a standard association binding entropy of less than 10 J/K*mol, ii) an absolute standard dissociation binding entropy of 100 J/mol*K or more, iii) an absolute standard binding entropy of 100 J/mol*K or more, e) producing an antibody by cultivating said selected cell under conditions suitable for the expression of said antibody and recovering said antibody from the cells or/and the cultivation medium.

Method to screen high affinity antibody

The present disclosure relates to an antibody that specifically binds a mutated NT-proBNP having i) a mutation substituting arginine at position 46 with histidine or ii) a mutation substituting glutamic acid at position 43 with aspartic acid. Moreover, the present disclosure relates to a mutated NT-proBNP or fragment thereof. Further, envisaged by the present disclosure are kits containing the antibody of the present disclosure, or the mutated NT-proBNP of the present disclosure. The present disclosure also concerns a method for diagnosing heart failure.

Antibodies recognizing genetic variants of NT-proBNP

The current invention is directed to the velocity factor. Based on the velocity factor antibodies can be classified, i.e. antibodies can be characterized on their binding properties as e.g. entropic or enthalpic antigen binder. A velocity factor based classification does not require detailed thermodynamic determinations and/or calculations. The velocity factor is the ratio of the antigen-antibody complex association rate constants ka determined at 37&#xb0; C. and 13&#xb0; C. As only two experimental determinations are required to calculate the velocity factor this is a fast and high-throughput suited method.

Velocity factor

The present invention concerns a reagent composition for releasing vitamin D compounds bound to vitamin D-binding protein, a method for the detection of a 25-hydroxyvitamin D compound in which the 25-hydroxyvitamin D compound is released from vitamin D-binding protein using this reagent and the mixture obtained in this manner is analyzed, the use of the reagent to release vitamin D compounds as well as a kit for detecting 25-hydroxyvitamin D which contains the reagent for releasing vitamin D compounds in addition to the usual immunological reagents.

Release reagent for vitamin D compounds

The current invention reports a method for producing an antibody comprising the steps of a) providing a plurality of hybridoma cells each expressing an antibody, b) determining the time dependent amount of said antibody bound to the respective antigen by surface plasmon resonance at different temperatures and different antibody concentrations, c) calculating with the time dependent amount determined in b) based on equations (II) to (XIII) at least the thermodynamic parameters (i) standard association binding entropy formula (A), (ii) standard dissociation binding entropy formula (B), (iii) standard binding entropy (&#x394;S&#xb0;), (iv) free standard binding enthalpy (&#x394;G&#xb0;), (v) standard dissociation free binding enthalpy formula (C), (vi) standard association free binding enthalpy formula (D), (vii) &#x2212;T&#x394;S&#xb0;, (viii) dissociation rate constant k, (ix) equilibrium binding constant K, and (x) association rate constant k, d) selecting a hybridoma cell producing an antibody with at least two of the following: i) a standard association binding entropy of less than 10 J/K*mol, ii) an absolute standard dissociation binding entropy of 100 J/mol*K or more, iii) an absolute standard binding entropy of 100 J/mol*K or more, e) producing an antibody by cultivating said selected cell under conditions suitable for the expression of said antibody and recovering said antibody from the cells or/and the cultivation medium.

The present invention relates to a method of measuring a vitamin D metabolite in a sample, the method comprising the steps of (a) treating said sample with a vitamin D metabolite releasing reagent under conditions appropriate to release a vitamin D metabolite from vitamin D-binding protein and not to cause protein precipitation, (b) subjecting the treated sample obtained in step (a) to a chromatographic separation, and (c) measuring a vitamin D metabolite during or after said chromatographic separation. The present invention also relates to methods for determining the vitamin D status of a subject, for use in the diagnosis of disease, and to agents and kits for use in performing the methods of the invention.

Measurement of vitamin D

Growing community of inventors

Hohenpeissenberg, Germany

Leopold Von Proff