公开(公告)号：US20240002443A1

公开(公告)日：2024-01-04

申请号：US18137604

申请日：2023-04-21

Applicant: BIOMADISON, INC

Inventor： Ward C Tucker ,  Francis Mark Dunning

IPC: C07K14/00 ,  C12Q1/37 ,  G01N33/542 ,  C12N9/52

CPC classification number: C07K14/00 ,  C12Q1/37 ,  G01N2333/33 ,  C12N9/52 ,  G01N33/542

Abstract: Recombinant nucleic acids encoding reporting constructs for characterizing botulinum neurotoxin protease activity and cells that incorporate such recombinant nucleic acids and that are suitable for use in cell-based assays for the neurotoxin are provided. The encoded reporting constructs are a pair of recombinant hybrid proteins that act in concert. The reporting constructs are a pair of recombinant hybrid proteins that act in concert, and that include a botulinum neurotoxin protease recognition and cleavage sequence positioned to release a fluorophore upon cleavage.

公开(公告)号：US20230143149A1

公开(公告)日：2023-05-11

申请号：US18094663

申请日：2023-01-09

Applicant: BioMadison, Inc.

Inventor： Francis Mark Dunning ,  Ward C. Tucker

IPC: C07K1/34 ,  G01N33/68 ,  B01D61/16 ,  B01D15/34 ,  B01D61/14

CPC classification number: C07K1/34 ,  G01N33/6803 ,  B01D61/16 ,  B01D15/34 ,  B01D61/145 ,  B01D2311/12

Abstract: Systems are described in which a primary detergent or surfactant in an aqueous solution is removed by the addition of a secondary detergent or surfactant in concentrations that exceed the critical micellar concentration (CMC) of the secondary detergent or surfactant using a size separation device. These systems are particularly applicable to protein-containing solutions. Typical primary detergents/surfactants include polysorbate 20, polysorbate 80, and Triton X-100. Suitable secondary detergents or surfactants can be ionic, nonionic, or zwitterionic. Typical secondary detergents/surfactants include, but are not limited to, galactoside detergents (e.g. octyl-β-galactoside), glucamide detergents (e.g. MEGA 8, MEGA 9, MEGA 10), cholamide detergents (e.g. CHAPS, CHAPSO, BIGCHAPS), and sulfobetaine detergents (such as sulfobetaine 3-10).

公开(公告)号：US11357838B2

公开(公告)日：2022-06-14

申请号：US16557450

申请日：2019-08-30

Applicant: BIOMADISON, INC.

Inventor： Francis Mark Dunning ,  Ward Tucker

IPC: A61K38/48 ,  C07K14/47 ,  C07K14/705 ,  C07K14/435 ,  C12N9/52 ,  C12Q1/37

Abstract: A protease directed to a non-neuronal SNARE protein is described. The protease is produced by selective mutation of a botulinum neurotoxin light chain, and is characterized utilizing a reporting construct that includes all or part of the non-neuronal SNARE protein. Such a protease has utility in the treatment of diseases associated with hypersecretion, where the hypersecretion is mediated by a non-neuronal SNARE protein.

公开(公告)号：US10526637B2

公开(公告)日：2020-01-07

申请号：US15980454

申请日：2018-05-15

Applicant: BIOMADISON, INC.

Inventor： Robert D. Fish ,  Min Dong

IPC: G01N31/00 ,  G01N33/53 ,  C12Q1/37 ,  G01N33/542 ,  G01N33/94 ,  C07K19/00 ,  C07K14/47 ,  C07K14/00 ,  G01N21/64

Abstract: Methods for improving the specificity of assays for botulinum neurotoxins are described. Reporting constructs are provided that include cleavable linker sequence with genetically modified recognition and/or cleavage sites. These linker sequences act as a substrate for a botulinum neurotoxin, with cleavage of the reporting construct providing a detectable signal. When first and second neurotoxins have activity with the same substrate protein, mutation and/or deletion of a recognition and/or cleavage site associated with the second neurotoxin improves the specificity of the detectable signal for the first neurotoxin.

公开(公告)号：US20190339273A1

公开(公告)日：2019-11-07

申请号：US16387389

申请日：2019-04-17

Applicant: BIOMADISON, INC.

Inventor： Ward C Tucker

IPC: G01N33/569 ,  C12N5/0781 ,  G01N33/50 ,  G01N33/94

Abstract: Compositions and methods for determining the efficacy and/or potency of a vaccine preparation are described herein. Splenocytes from immunized animals are isolated and frozen. Upon thawing aliquots these cells are activated by exposure to a series of dilutions of q vaccine preparation being tested and a series of dilutions of a reference vaccine with known characteristics. Cells secreting immunogen-specific antibody and cells secreting nonspecific antibody are enumerated, as is the amount of immunogen-specific and nonspecific antibody produced. Comparison between the results from the vaccine preparations provides a measure of relative vaccine efficacy and/or potency.

公开(公告)号：US20190331665A1

公开(公告)日：2019-10-31

申请号：US16408308

申请日：2019-05-09

Applicant: BIOMADISON, INC.

Inventor： Tim Piazza ,  Ward C Tucker

IPC: G01N33/50 ,  G01N33/58 ,  G01N33/569 ,  C12Q1/37

Abstract: Compositions and methods are provided that improve detection of botulinum neurotoxins in cell-based assays. In one aspect an isoquinolynyl compound can be used to enhance the sensitivity of both Förster resonance energy transfer (FRET) and non-FRET cell-based assays. Osmolarity of the cell culture media can be adjusted to optimize the effect of the compound. In that subject matter an environment cell can include an enzyme that facilitates degradation of the reporter significantly faster after the cleavage than before the cleavage, and presence of the Botulinum toxin correlates with reduction of the signal from a baseline signal. Where the environment is a cell, the cell can advantageously express both the construct that includes the reporter, and an enzyme that facilitates the degradation.

公开(公告)号：US09526345B2

公开(公告)日：2016-12-27

申请号：US14455786

申请日：2014-08-08

Applicant: BIOMADISON, INC.

Inventor： Ward C. Tucker ,  Tim Piazza

IPC: C12Q1/37 ,  A47C15/00 ,  G01N21/64 ,  G01N33/569 ,  C12Q1/68

CPC classification number: G01N33/56911 ,  A47C15/006 ,  C12Q1/37 ,  C12Q1/6897 ,  G01N21/6408 ,  G01N21/6428 ,  G01N2333/33

Abstract: Methods and compositions are provided where a transfected cell that produces a hybrid protein with a reporter-containing portion and a botulinum toxin cleavage site is contacted with a botulinum toxin at elevated temperatures and/or in media having a reduced sodium concentration. Kits that include such media and a botulinum toxin are also described.

Abstract translation: 提供了方法和组合物，其中产生具有报道分子部分和肉毒杆菌毒素切割位点的杂交蛋白的转染细胞在升高的温度和/或具有降低的钠浓度的培养基中与肉毒杆菌毒素接触。 还描述了包括这种介质和肉毒杆菌毒素的试剂盒。

公开(公告)号：US11685945B2

公开(公告)日：2023-06-27

申请号：US16784121

申请日：2020-02-06

Applicant: BIOMADISON, INC.

Inventor： Ward C Tucker

IPC: C12N15/85 ,  C12Q1/37 ,  C07K14/435

CPC classification number: C12Q1/37 ,  C07K14/43595 ,  C07K2319/03 ,  C07K2319/50 ,  C07K2319/60 ,  G01N2333/952

Abstract: Compositions and methods are disclosed that provide a rapid, sensitive, and accurate cell-based assay for enzyme activity, particularly for enzyme activities associated with botulinum toxins. A cell is provided that expresses a construct that includes an anchor region, a cleavage site, and a reporting region having two or more identical reporter peptides. Enzymatic activity at the cleavage site releases the reporter region into the cytosol, where multiple degradation events occur. The observed change in the signal is proportional to the enzymatic activity.

公开(公告)号：US11661442B2

公开(公告)日：2023-05-30

申请号：US17208649

申请日：2021-03-22

Applicant: BIOMADISON, INC

Inventor： Ward C Tucker ,  Francis Mark Dunning

IPC: C07K14/00 ,  C12Q1/37 ,  G01N33/542 ,  C12N9/52

CPC classification number: C07K14/00 ,  C12N9/52 ,  C12Q1/37 ,  G01N33/542 ,  G01N2333/33

Abstract: Vesicles that incorporate reporting constructs for characterizing Botulinum neurotoxin protease activity and suitable for use in an assay are provided. The reporting constructs are a pair of recombinant hybrid proteins that act in concert. The reporting constructs are a pair of recombinant hybrid proteins that act in concert, and that include a Botulinum neurotoxin protease recognition and cleavage sequence positioned to release a fluorophore upon cleavage.

公开(公告)号：US20200291065A1

公开(公告)日：2020-09-17

申请号：US16818678

申请日：2020-03-13

Applicant: BIOMADISON, INC.

Inventor： Francis Mark DUNNING ,  Ward C. Tucker

IPC: C07K1/34 ,  G01N33/68 ,  B01D61/14 ,  B01D61/16 ,  B01D15/34

Abstract: Compositions and methods are described in which a primary detergent or surfactant in an aqueous solution is removed by the addition of a secondary detergent or surfactant in concentrations that exceed the critical micellar concentration (CMC) of the secondary detergent or surfactant. These compositions and methods are particularly applicable to protein-containing solutions. Typical primary detergents/surfactants include polysorbate 20, polysorbate 80, and Triton X-100. Suitable secondary detergents or surfactants can be ionic, nonionic, or zwitterionic. Typical secondary detergents/surfactants include, but are not limited to, galactoside detergents (e.g. octyl-β-galactoside), glucamide detergents (e.g. MEGA 8, MEGA 9, MEGA 10), cholamide detergents (e.g. CHAPS, CHAPSO, BIGCHAPS), and sulfobetaine detergents (such as sulfobetaine 3-10).