Title:
EXTERNAL GRADIENT CHROMATOFOCUSING
Document Type and Number:
WIPO Patent Application WO2004103519
Kind Code:
A3
Abstract:
The invention provides novel methods for the separation of charged molecules such a proteins according to their isoelectric points (pI's) and includes the systems and buffering compositions employed for isolating charged molecules. The invention further provides for modifications to the above described chromatographic methods that enable the separation of charged molecules exhibiting virtually identical pl's by shifting both the buffer's pKa and the pl's of the eluted charged molecules while they are traversing the ion exchange column.
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Inventors:
TSONEV LATCHEZAR I (US)
Application Number:
PCT/US2004/015216
Publication Date:
February 03, 2005
Filing Date:
May 14, 2004
Export Citation:
Assignee:
DNAVAX INC (US)
TSONEV LATCHEZAR I (US)
TSONEV LATCHEZAR I (US)
International Classes:
B01D15/16; B01D15/36; C07K1/16; C07K1/18; C07K1/28; G01N30/02; (IPC1-7): B01D15/08; C07K1/18; G01N30/60
Foreign References:
| EP0172217A1 | 1986-02-26 | |||
| EP1273592A2 | 2003-01-08 |
Other References:
YANSHENG L ET AL: "Gradient chromatofocusing high-performance liquid chromatography I. Practical aspects", JOURNAL OF CHROMATOGRAPHY A, ELSEVIER SCIENCE, NL, vol. 762, no. 1-2, 21 February 1997 (1997-02-21), pages 207 - 217, XP004116706, ISSN: 0021-9673
WALTHER-RASMUSSEN J ET AL: "Hydrophobic character of surface regions and total hydrophobicity of four variants of chromosomal class C beta-lactamase from Pseudomonas aeruginosa are identical. - Chromatographic comparison of the hydrophobic character of the variants and the effect of focusing buffer composition on the separatio", JOURNAL OF CHROMATOGRAPHY. BIOMEDICAL APPLICATIONS, ELSEVIER, AMSTERDAM, NL, vol. 746, no. 2, 15 September 2000 (2000-09-15), pages 161 - 172, XP004210422, ISSN: 0378-4347
MHATRE R ET AL: "Purification of antibody Fab fragments by cation-exchange chromatography and pH gradient elution", JOURNAL OF CHROMATOGRAPHY A, ELSEVIER SCIENCE, NL, vol. 707, no. 2, 21 July 1995 (1995-07-21), pages 225 - 231, XP004038812, ISSN: 0021-9673
DATABASE COMPENDEX [online] ENGINEERING INFORMATION, INC., NEW YORK, NY, US; 20 January 1999 (1999-01-20), LOGAN KATHLEEN A ET AL: "Simple, two-component buffer enhances use of chromatofocusing for processing of therapeutic proteins", XP002294452, Database accession no. EIX99174584614
KANG X ET AL: "High-performance cation-exchange chromatofocusing of proteins", JOURNAL OF CHROMATOGRAPHY A, ELSEVIER SCIENCE, NL, vol. 991, no. 1, 28 March 2003 (2003-03-28), pages 117 - 128, XP004413645, ISSN: 0021-9673
See also references of EP 1628729A2
WALTHER-RASMUSSEN J ET AL: "Hydrophobic character of surface regions and total hydrophobicity of four variants of chromosomal class C beta-lactamase from Pseudomonas aeruginosa are identical. - Chromatographic comparison of the hydrophobic character of the variants and the effect of focusing buffer composition on the separatio", JOURNAL OF CHROMATOGRAPHY. BIOMEDICAL APPLICATIONS, ELSEVIER, AMSTERDAM, NL, vol. 746, no. 2, 15 September 2000 (2000-09-15), pages 161 - 172, XP004210422, ISSN: 0378-4347
MHATRE R ET AL: "Purification of antibody Fab fragments by cation-exchange chromatography and pH gradient elution", JOURNAL OF CHROMATOGRAPHY A, ELSEVIER SCIENCE, NL, vol. 707, no. 2, 21 July 1995 (1995-07-21), pages 225 - 231, XP004038812, ISSN: 0021-9673
DATABASE COMPENDEX [online] ENGINEERING INFORMATION, INC., NEW YORK, NY, US; 20 January 1999 (1999-01-20), LOGAN KATHLEEN A ET AL: "Simple, two-component buffer enhances use of chromatofocusing for processing of therapeutic proteins", XP002294452, Database accession no. EIX99174584614
KANG X ET AL: "High-performance cation-exchange chromatofocusing of proteins", JOURNAL OF CHROMATOGRAPHY A, ELSEVIER SCIENCE, NL, vol. 991, no. 1, 28 March 2003 (2003-03-28), pages 117 - 128, XP004413645, ISSN: 0021-9673
See also references of EP 1628729A2
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