Oxford Congenital Myasthenia Detection Service Genetic testing
Structural and useful characterization of C0021158, a high-affinity monoclonal antibody that inhibits Arginase
IvanSeptember 8, 20200 Comments
Structural and useful characterization of C0021158, a high-affinity monoclonalantibody that inhibits Arginase 2 perform by way of a novel non-competitive mechanism of motion
Arginase 2 (ARG2) is a binuclear manganese metalloenzyme that catalyzes the hydrolysis of L-arginine. The dysregulated expression of ARG2 inside particular tumor microenvironments generates an immunosuppressive area of interest that successfully renders the tumor ‘invisible’ to the host’s immune system. Elevated ARG2 expression results in a concomitant depletion of native L-arginine ranges, which in flip results in suppression of anti-tumor T-cell-mediated immune responses.
Right here we describe the isolation and characterization of a excessive affinity antibody (C0021158) that inhibits ARG2 enzymatic perform fully, successfully restoring T-cell proliferationin vitro. Enzyme kinetic research confirmed that C0021158 displays a noncompetitive mechanism of motion, inhibiting ARG2 independently of L-arginine concentrations.
To elucidate C0021158’s inhibitory mechanism at a structural degree, the co-crystal construction of the Fab in advanced with trimeric ARG2 was solved. C0021158’s epitope was consequently mapped to an space far from the enzyme’s substrate binding cleft, indicating an allosteric mechanism was being employed. Following C0021158 binding, distinct areas of ARG2 bear main conformational modifications.
Notably, the spine construction of a surface-exposed loop is totally rearranged, resulting in the formation of a brand new quick helix construction on the Fab-ARG2 interface. Furthermore, this large-scale structural reworking at ARG2’s epitope interprets into extra delicate modifications inside the enzyme’s lively web site. An arginine residue at place 39 is reoriented inwards, sterically impeding the binding of L-arginine. Arg39 can also be predicted to change the pOkA of a key catalytic histidine residue at place 160, additional attenuating ARG2’s enzymatic perform.
In silico molecular docking simulations predict that L-arginine is unable to bind successfully when antibody is certain, a prediction supported by isothermal calorimetry experiments utilizing an L-arginine mimetic. Particularly, concentrating on ARG2 within the tumor microenvironment by means of the applying of C0021158, probably together with commonplace chemotherapy regimens or alternate immunotherapies, represents a possible new technique to focus on immune chilly tumors.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Mouse Pepsin (PP) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Mouse Pepsin (PP) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Mouse Pepsin (PP) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Mouse Pepsin (PP) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: Enzyme-linked immunosorbent assay based on the Competitive Inhibition method for detection of Mouse Pepsin (PP) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids with no significant corss-reactivity with analogues from other species.
Formulating monoclonalantibodies as powders for reconstitution at excessive focus utilizing spray-drying: Trehalose/amino acid combos as reconstitution time decreasing and stability enhancing formulations
To extend their stability, therapeutic (or monoclonal) antibodies (mAbs) are sometimes formulated as solids by utilizing quite a lot of drying methods, e.g. freeze-drying, spray-drying, or spray freeze-drying. The addition of excipients is required to protect stability of the protein in the course of the drying course of and subsequent storage of the ensuing strong type. The addition of low molecular weight excipients, similar to amino acids, to sugar based mostly spray- and freeze-dried formulations has been recommended to enhance the storage stability of proteins within the dried state.
On this examine sugars (sucrose, trehalose), amino acids (Gly, Ala, Professional, Ser, Val, Leu, Ile, Gln, His, Lys, Arg, Phe, Trp) and combos thereof have been investigated for his or her stabilizing impact throughout spray-drying and subsequent storage and for his or her reconstitution time decreasing impact.
Two IgG4 mAbs have been used as mannequin antibodies. From an preliminary screening examine, fundamental and small impartial amino acids, together with a sugar, similar to sucrose or trehalose, confirmed reconstitution time decreasing and stabilizing properties. Arg particularly displayed glorious reconstitution and stability enhancing properties. Furthermore, Arg was the one amino acid offering stabilizing properties akin to sucrose or trehalose.
Earlier work by the authors described a statistically substantiated comparability between the three fundamental amino acids in a sugar containing formulation, albeit restricted to a single focus degree [5]. Due to this fact, a follow-up design of experiments (DoE) examine was carried out to find out the optimum trehalose/amino acid content material required for an optimum protein stability and reconstitution time and to check the results of two fundamental amino acids, Lys and Arg, to these of two impartial amino acids, Gly and Professional. The performed DoE coated a variety of trehalose (30 – 120 mM) and amino acid (50 – 150 mM) concentrations.
The focus of trehalose was discovered to be the principle contributor to a discount in reconstitution time and a rise in stability. Right here we present that the addition of amino acids similar to Gly, Professional, and Lys doesn’t enhance stability nor does it scale back the reconstitution time. Of the examined amino acids, solely Arg confirmed a marked discount in reconstitution time and enchancment in stability in comparison with a trehalose.
Furthermore, the properties displayed by Arg might justify its software as the principle stabilizer in spray-dried mAb formulations, eliminating the necessity for a sugar matrix altogether. However the weight ratio of stabilizer to protein was discovered the issue exerting the strongest general affect on the formulation’s reconstitution time and stability. Extra particularly, adequate bodily stability and an appropriate reconstitution time might be obtained with a protein to stabilizer weight ratio of not less than 1:1.
Description: A Monoclonal antibody against Human GHRL / Ghrelin (clone 2F4). The antibodies are raised in Mouse and are from clone 2F4. This antibody is applicable in WB and IHC-P, E
Description: A Monoclonal antibody against Human GHRL / Ghrelin (clone 4B8). The antibodies are raised in Mouse and are from clone 4B8. This antibody is applicable in WB and IHC-P, E
Description: Ghrelin is the ligand for growth hormone secretagogue receptor type 1 (GHSR). Induces the release of growth hormone from the pituitary. Has an appetite-stimulating effect, induces adiposity and stimulates gastric acid secretion. Involved in growth regulation.
Description: This gene encodes the ghrelin-obestatin preproprotein that is cleaved to yield two peptides, ghrelin and obestatin. Ghrelin is a powerful appetite stimulant and plays an important role in energy homeostasis. Its secretion is initiated when the stomach is empty, whereupon it binds to the growth hormone secretagogue receptor in the hypothalamus which results in the secretion of growth hormone (somatotropin). Ghrelin is thought to regulate multiple activities, including hunger, reward perception via the mesolimbic pathway, gastric acid secretion, gastrointestinal motility, and pancreatic glucose-stimulated insulin secretion. It was initially proposed that obestatin plays an opposing role to ghrelin by promoting satiety and thus decreasing food intake, but this action is still debated. Recent reports suggest multiple metabolic roles for obestatin, including regulating adipocyte function and glucose metabolism. Alternative splicing results in multiple transcript variants. In addition, antisense transcripts for this gene have been identified and may potentially regulate ghrelin-obestatin preproprotein expression.,Amino acids QRKESKKPPAKLQPR from the human protein were used as the immunogen for the Ghrelin antibody.,After reconstitution
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Mouse Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Mouse Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Mouse Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Mouse Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: Enzyme-linked immunosorbent assay based on the Competitive Inhibition method for detection of Mouse Ghrelin (GHRL) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids with no significant corss-reactivity with analogues from other species.
Description: Quantitativesandwich ELISA kit for measuring Mouse ghrelin (GHRL) in samples from serum, plasma, cell culture supernates, tissue homogenates. A new trial version of the kit, which allows you to test the kit in your application at a reasonable price.
Description: Quantitativesandwich ELISA kit for measuring Mouse ghrelin (GHRL) in samples from serum, plasma, cell culture supernates, tissue homogenates. Now available in a cost efficient pack of 5 plates of 96 wells each, conveniently packed along with the other reagents in 5 separate kits.
Description: A sandwich quantitative ELISA assay kit for detection of Mouse Ghrelin (GHRL) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Mouse Ghrelin (GHRL) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: Enzyme-linked immunosorbent assay based on the Double-antibody Sandwich method for detection of Mouse Ghrelin (GHRL) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids with no significant corss-reactivity with analogues from other species.
Description: A polyclonal antibody raised in Goat that recognizes and binds to Human GHRL / Ghrelin (C-Terminus). This antibody is tested and proven to work in the following applications:
Description: A sandwich ELISA kit for detection of Ghrelin from Mouse in samples from blood, serum, plasma, cell culture fluid and other biological fluids.
Description: A competitive Inhibition ELISA kit for detection of Ghrelin from Mouse in samples from blood, serum, plasma, cell culture fluid and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mini Samples Mouse Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mini Samples Mouse Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mini Samples Mouse Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mini Samples Mouse Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: Enzyme-linked immunosorbent assay based on the Double-antibody Sandwich method for detection of Mini Samples Mouse Ghrelin (GHRL) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids with no significant corss-reactivity with analogues from other species.
Description: A Monoclonal antibody against Human GHRL (monoclonal) (M01). The antibodies are raised in mouse and are from clone 2F4. This antibody is applicable in WB and IHC, E
Description: A Monoclonal antibody against Human GHRL (monoclonal) (M09). The antibodies are raised in mouse and are from clone 4B8. This antibody is applicable in WB and IHC, E
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Pig Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Pig Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Pig Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Pig Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: Enzyme-linked immunosorbent assay based on the Competitive Inhibition method for detection of Pig Ghrelin (GHRL) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids with no significant corss-reactivity with analogues from other species.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Rat Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Rat Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Rat Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Competitive Enzyme-linked immunosorbent assay for detection of Rat Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: Enzyme-linked immunosorbent assay based on the Competitive Inhibition method for detection of Rat Ghrelin (GHRL) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids with no significant corss-reactivity with analogues from other species.
Description: Quantitativesandwich ELISA kit for measuring Rat ghrelin (GHRL) in samples from serum, plasma, cell culture supernates. A new trial version of the kit, which allows you to test the kit in your application at a reasonable price.
Description: Quantitativesandwich ELISA kit for measuring Rat ghrelin (GHRL) in samples from serum, plasma, cell culture supernates. Now available in a cost efficient pack of 5 plates of 96 wells each, conveniently packed along with the other reagents in 5 separate kits.
Description: A competitive inhibition quantitative ELISA assay kit for detection of Rat Ghrelin (GHRL) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A competitive inhibition quantitative ELISA assay kit for detection of Rat Ghrelin (GHRL) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: Quantitativesandwich ELISA kit for measuring Pig Ghrelin (GHRL) in samples from serum, plasma, tissue homogenates. A new trial version of the kit, which allows you to test the kit in your application at a reasonable price.
Description: Quantitativesandwich ELISA kit for measuring Pig Ghrelin (GHRL) in samples from serum, plasma, tissue homogenates. Now available in a cost efficient pack of 5 plates of 96 wells each, conveniently packed along with the other reagents in 5 separate kits.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Rat Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Rat Ghrelin (GHRL) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
