Chemistry: analytical and immunological testing – Involving an insoluble carrier for immobilizing immunochemicals
Reexamination Certificate
2000-08-15
2003-04-29
Le, Long V. (Department: 1641)
Chemistry: analytical and immunological testing
Involving an insoluble carrier for immobilizing immunochemicals
C436S008000, C436S085000, C436S161000, C436S174000, C436S176000, C436S178000, C436S518000, C436S523000, C436S524000, C436S528000, C436S529000, C436S530000, C436S531000, C436S532000, C436S533000, C436S541000, C436S823000, C436S824000, C435S004000, C435S006120, C435S007100, C435S007900, C435S174000, C435S176000, C435S177000, C435S178000, C435S179000, C435S180000, C435S181000, C435S183000, C435S803000, C435S814000, C435S963000, C422S040000
Reexamination Certificate
active
06555391
ABSTRACT:
CROSS-REFERENCES TO RELATED APPLICATIONS
Not applicable.
BACKGROUND OF THE INVENTION
Affinity chromatography is a powerful method used to purify molecules and pharmaceutical compounds. A wide variety of molecules have been purified using various affinity purification schemes. Generally, the synthesis of affinity chromatography supports involves coupling a ligand having an affinity for the molecule being purified to a chromatography support. Through the specific interactions between the molecule and its ligand, significant increases in the purity of the molecule over the starting material can be achieved or significant amounts of a molecule can be removed from a starting material.
Typically, the synthesis of affinity chromatography resins involves reacting a ligand with a functional group on a chromatographic support or a derivatized group on a chromatographic support to form a ligand-matrix bond. Unfortunately, not all ligand-chromatographic support bonds are stable and the chromatographic support itself may trap ligands (or complexes of ligands) that are not bound to a functional group. Furthermore, these ligands can leach from the column during purification, contaminating the product. This is often a serious concern, especially in the pharmaceutical arena. Often regulatory agencies will issue strict guidelines to pharmaceutical companies for the amount of ligand that can be present in a pharmaceutical composition; during the regulatory approval process and on approved drugs, regulatory agencies (e.g., the FDA in the US) require that stringent limits (or specifications) be met on the amounts of contaminants such as ligands that can be present in the pharmaceutical. Thus, the presence of these contaminants can result in a ruined batch of the pharmaceutical or in increased expenditures of time and effort to further purify the pharmaceutical away from the ligand. Moreover, immunization of patients against the drug or anaphylactic shock could result if the ligand contaminant is an antibody and the molecule is used in a clinical setting (Ubrich and Rivat (1996)
Art. Cells, Blood Subs., and Immob. Biotech
. 24: 65-75). Therefore a need in the art exists for methods of decreasing the amount of ligand that leaches from an affinity column during purification of the molecule of choice. The present invention fulfills these and other needs.
SUMMARY OF THE INVENTION
The present invention provides methods for conditioning a chromatography resin after storage. The methods involve contacting a chromatography resin with an effective amount of a hydroxyalkyl amine compound that comprises at least one hydroxyl moiety. An example of a suitable compound is ethanolamine. The methods are generally used on a chromatography resin that is composed of a ligand that was conjugated to a chromatographic support at least 48 hours prior to contacting the chromatography resin with the hydroxyalkyl amine compound. For example, the invention provides methods for conditioning a chromatography resin that consists of an activated hydroxyl chromatographic support to which is attached an affinity ligand.
The invention also encompasses methods for preparing an affinity resin from which less ligand will leach during purification of a molecule to which the ligand binds. The methods entail contacting an affinity resin with a hydroxyalkyl amine compound. The affinity resin is composed of a ligand that is conjugated to a chromatographic support, and has been stored for at least 48 hours after coupling the ligand to the chromatographic support. The affinity ligand is incubated in the presence of the hydroxyalkyl amine compound for a time sufficient to remove ligand that is not stably bound to the chromatography resin. Sufficient unstably bound ligand is removed by the treatment so that when the treated affinity resin is used for affinity purification of a molecule to which the ligand specifically binds, less ligand leaches from the affinity resin as compared to a purification of the molecule on the affinity resin that has not been treated with the compound.
REFERENCES:
patent: 5126412 (1992-06-01), Das et al.
patent: 5525472 (1996-06-01), Lifshitz et al.
patent: 6150151 (2000-11-01), Khandke
Tharakan et al. Physical and biochemical characterization of five commercial resins for immunoaffinity purification of Factor IX. Journal of Chromatography. (1992) vol. 595, Nos. 1-2, pp. 103-111.*
Highsmith et al. Evaluation of cyanogen bromide, FMP and hydrazide resins for immunoaffinity purification of Factor IX. Biotechniques. (1992) vol. 12, No. 3, pp. 418, 420, 422-423.*
Harlow and Lane: “Antibodies—A Laboratory Manual”Cold Springs Harbor Laboratory Publications, N.Y.(1988).
Ubrich and Rivat: “Antibodies Released From Immunoadsorbents: Effect of Support, Activation and Elution Conditions”Art, Cells, Blood Subs., and Immob. Biotech.24:65-75 (1996).
Antonsen et al.: “Elution Conditions and Degradation Mechanisms in Long-Term Immunoadsorbent Use”Biotechnol. Prog.7:159-172 (1991).
van Sommeren et al.: “Comparison of three activated agaroses for use in affinity chromatography: effects on coupling performance and ligand leakage”J. Chromatrography639:23-31 (1993).
Scopes, R.K.: “Protein Purification, Principles and Practice” pp. 111-135,Springer-Verlag New York Inc.(1982).
Kohn and Wilchek: “The Use of Cyanogen Bromide and other Novel Cyanylating Agents for the Activation of Polysaccharide Resins”Applied Biochem. and Biotechnol.9:285-305 (1984).
March et al.: “A Simplifed Method for Cyanogen Bromide Activation of Agarose for Affinity Chromatography”Anal. Biochem.60:149-152 (1974).
Jennissen, H.P.: “Cyanogen Bromide and Tresyl Chloride Chemistry Revisted: The Special Reactivity of Agarose as a Chromatographic and Biomaterial Support for Immobilizing Novel Chemical Groups”J. Mol. Recog.8:116-124 (1995).
Desai, M.A.: “Immunoaffinity Adsorption: Process-Scale Isolation of Therapeutic-Grade Biochemicals”J. Chem. Techn. Biotechnol.48:105-126 (1990).
Berry and Pierce: “Stability of immunoadsorbents comprising antibody fragments. Comparison of Fv fragments and single-chain Fv fragments”J. Chromatog.629:161-168 (1993).
Wilchek et al.: “Structure of a Soluble Super-Active Insulin Is Revealed by the Nature of the Complex Between Cyanogen-Bromide-Activated Sepharose and Amines”Proc. Nat. Sci. USA72:1055-1058 (1975).
Bernhard Susan L.
Taiariol Van
Toso Robert
Baxter International Inc.
Le Long V.
Padmanabhan Kartic
Townsend and Townsend / and Crew LLP
LandOfFree
Methods of conditioning an affinity resin after storage does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Methods of conditioning an affinity resin after storage, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Methods of conditioning an affinity resin after storage will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3094255