PolyCAT A

PolyCAT A

- for cation-exchange of proteins

PolyCAT A is made through a unique process for attaching poly(aspartic acid) covalently to silica. Proteins elute from this polypeptide coating in sharp peaks with little tailing. Binding capacity and recovery are high as well.Operating conditions are similar to those used with other weak cation-exchange (WCX-) materials (eg. CM-type).

When to use PolyCAT A:

  1. Hemoglobin analysis (Hb A1c and variants)
  2. Proteins with isoelectric points above 6.0.
  3. Polypeptides with more than 3 basic residues, such as hormones and growth factors (easier elution than with PolySULFOETHYL A).
  4. Purification of very basic solutes by ion-exchange with a volatile mobile phase.
  5. Quality control assay of isoforms, cf. sialylation variants of monoclonal antibodies.
  6. Histones (using mixed modes of ion-exchange and hydrophilic interaction).

With larger proteins such as hemoglobins and immunoglobulins, selectivity is optimal with PolyCAT A with 1000-angstrom pores.

Tips on Ion-exchange:

  1. Volatile mobile phases

    2. It is possible to use our ion-exchange columns with volatile mobile phases. Peptides are adsorbed from dilute solutions of ammonium acetate and eluted with a step or linear gradient to dilute (@5-10%) acetic acid. With very basic peptides, use PolyCAT A; otherwise, use PolySULFOETHYL A. This method also works with other very basic solutes, such as aminoglycoside antibiotics.  Ask for our applications sheet about this.

  2. Organic solvents in the mobile phases

    3. When the mobile phase contains over 40% organic solvent, then our cation-exchangers become sensitive to changes in polar as well as basic residues. Examples include the presence or absence of a Ser-residue or the methylation of a Lys-residue. This is proving to be particularly useful for resolution of minor forms of large recombinant or synthetic  polypeptides.                                                                                                            

Paper  on PolyCAT A

Subject

C1.  Alpert, J. Chromatogr. 266 (1983) 23.

Description  of PolyCAT A.

C2.  Ou et al., J. Chromatogr. 266 (1983) 197

Analysis  of hemoglobin variants

C3.  Parente & Wetlaufer, J. Chromatogr. 288, (1984) 389

Effect  of urea on cation-exchange of lysozyme

C4.  Aswad, J. Biol. Chem. 259 (1984) 10714

Cation-exchange  of ACTH

C5. Parente & Wetlaufer, J. Chromatogr. 314 (1984)  337

Effect of heat and urea on cation exchange of  chymotrypsinogen

C6.  Ellis et al., Clin. Chem. 30 (1984) 1746

Assay  of Hemoglobin A1c

C7.  Rogers et al., Am. J. Clin. Pathol. 84 (1985) 671

Analysis  of hemoglobinopathies

C8.  Ou & Carmona, Clin. Chem. 31 (1985) 945

Assay  of hemoglobin A1c (abstract only).

C9.  Parente & Wetlaufer, J. Chromatogr. 355,(1986) 29

Isocratic  vs. gradient elution of proteins

C10.  Wessels et al., Clin. Chem. 32 (1986) 903

Diagnosis  of Hemoglobin S/Lepore

C11.  Turpeinen, Clin. Chem. 32 (1986) 999

Assay  of Hemoglobin A2

C12.  Langer et al., Anal. Biochem. 166 (1987) 212

Purification  of N-Glycosidase F

C13.  Spiker et al., J. Chromatogr. 405 (1987) 327

Fractionation  of HMG proteins

C14.  Bisse & Wieland, J. Chromatogr. 434 (1988) 95

Assay  of Hemoglobins F and A1c

C15.  Kavanaugh et al., J. Biol. Chem. 264 (1989) 11009

Purif.  of ATP-labeled hemoglobin

C16.  Marchese et al., J. Chromatogr. 504 (1990) 351

Separation  of three forms of rh Interleukin-2

C17.  Canova-Davis et al., J. Chromatogr. 508 (1990) 81

Characterization  of synthetic human relaxin

C18.  Ebert & Spryn, Bioconjugate Chem. 1 (1990) 331

Purification  of a ribosome-inactivating protein from barley

C19.  Osheroff et al., J. Biol. Chem. 265 (1990) 9386

Prepn.  of 32P-labeled human relaxin

C20.  Kutlar et al., Hum. Genet. 85 (1990) 106

Assay  and purif. of Hemoglobin F

C21.  Rao et al., BioPharm 4 (#10) (1991) 38

Separation  of post-translational variants of monoclonal antibodies

C22.  Huisman, Sangre 36 (1991) 144

HPLC  of abnormal hemoglobins

C23.  Lee et al., Diabetes Res. Clin. Pract. 16 (1992) 53

HPLC  of glycated proteins vs. home blood glucose testing

C24.  Holmes et al., Science 256 (1992) 1205

Isolation  of heregulin (an oncogene product activator)

C25.  Burton et al., J. Neurochem. 59 (1992) 1937

Analysis  of various forms of rNerve Growth Factor

C26.  Fu et al., J. Biol. Chem. 266 (1991) 14562

Affinity-style  isoln. of an L-isoaspartyl Protein Carboxyl Methyltransferase

C27.  Van der Dijs et al., Clin. Chem. 38 (1992) 1864

Screening  cord blood for hemoglobino pathies and thalassemia.

C28.  Ou & Rognerud, Clin. Chem. 39 (1993) 820

Rapid  screening for hemoglobin variants.

C29.  Kutlar & Huisman, J. Chromatogr. 620 (1993) 183

Micro  HPLC method for determining the gamma-chain composition of hemoglobin  F

C30.  Hazelett et al., Alcohol. Clin. Exp. Res. 17(1993) 1107

Screening Hemoglobin-acetaldehyde as a marker for alcoholism

C31.  Gibbs et al., Proc. Natl. Acad. Sci. 88 (1991) 4001

Purif.  of single-chain catalytic antibody

C32.  Pantoliano et al., Biochemistry 30 (1991) 10117

Purif.  of single-chain anti-fluorescein antibody

C33.  Milenic et al., Cancer Res. 51 (1991) 6363

Purif.  of single-chain anti-carcinoma antibody

C34.  Stewart et al., Biochemistry 33 (1994) 1994

Purification  of catalytic antibody

C35.  Richard et al., Amer. Clin. Lab. (1/1994) 26

Assay  of Hemoglobin A1c

C36.  Lu et al., Chromosome 103 (1994) 111

Separation  of Histone H1phosphorylation variants from Tetrahymena

C37.  Itälä et al., Anal. Biochem. 224 (1995) 323

Hemoglobin-acetaldehyde analysis

C38.  Fiordalisi et al., Protein Exp. Purif. 3 (1992) 282

Purif.  of recombinant Bungarotoxin

C39.  Pearson et al., Hemoglobin 18 (1994) 401

HPLC  of rat hemoglobins

C40.  Thomas et al., Science 241 (1988) 226

Analysis  of prohormone versions of beta-endorphin and gamma lipotropin

C41.  Thorne et al., J. Biol. Chem. 264 (1989) 3545

Further  study of prohormone processing

C42.  Thomas et al., J. Biol. Chem. 265 (1990) 10821

More  prohormone processing

C43.  Thomas et al., Proc. Natl. Acad. Sci. 88 (1991) 5297

Still  more prohormone processing

C44.  Pic et al., Ann. Biol. Clin. 52 (1994) 129

Assay  of Hemoglobin F, Fac, & variants.

C45.  Jones, Meth. Enzymol. 231 (1994) 322

Chemically-modified hemoglobins

C46.  Turpeinen et al., Clin. Chem. 41 (1995) 191

Assay  of Hemoglobin A1c ; comparison of PolyCAT A with  Diamat  and affinity methods

C47.  Turpeinen et al., Clin. Chem. 41 (1995) 532

Assay  of Hemoglobin alpha-chain variants : Broussais & Cemenelum.

C48.  Bissé et al., Clin. Chem. 41 (1995) 658

Elevation  of Hemoglobin A1d in uremia

C49.  Cuevas et al., Clin. Chem. 41 (1995) S98 (abstract 292).

Retention  of Hemoglobin variants relative to the Hb A peak.

C50.  Ou & Rognerud, ibid, S100 (abstract 300).

Assay  of Hemoglobins Chicago & South Florida

C51.  Ou & Rognerud, Clin. Chem. 42 (1996) 774

Assay  of Hemoglobin Chicago/S trait and Hb Chicago/Sickle Cell  disease.

C52.  Shahrokh et al., Pharm. Res. 11 (1994) 936

Basic  Fibroblast Growth Factor. Isolation of an Asp15-succinimide15  variant and misc. cleavage variants.

C53.  Bissé et al., J. Chromatogr. B687 (1996) 349.

Elevation  of Hemoglobins A1d3a and A1d3b in uremia and diabetes,  resp.

C54.  Vunnam et al., J. Peptide Res. 49 (1997) 59.

Purif.   of synthetic cecropin and Pro-Arg-rich peptides from pig intestine

C55.  Chen et al., Alcohol Clin. Exp. Res. 19 (1995) 939.

New Hemoglobin-acetaldehyde variants

C56.  Wilson et al., J. Lab. Clin. Med. 108 (1986)138.

Analysis  of hemoglobins S, C, and O Arab in cord blood samples.

C57.  Fishman et al., J. Chromatogr. 532 (1990) 55

Separation  of pyridoxylated hemoglobins

C58.  Galacteros et al., Blood 78 (1991) 1121.

Adult hemoglobin conc. vs. gestational age in newborns.

C59.  Roa et al., Ann. Clin. Lab. Sci. 23 (1993) 433

Hemoglobin  variant detection in dried blood specimens

C60.  Roa et al., Ann. Clin. Lab. Sci. 25 (1995) 228

Reference  ranges for hemoglobin variants in African Americans.

C61.  Garibay-Tupas et al., J. Endocrinol. 145 (1995) 441

Prepn.  of 32P-labeled human relaxin

C62.  Huber et al., Prenat. Diagn. 16 (1996) 1011

Estimating hemoglobin F from fetal cells isolated from maternal  blood.

C63.  Leonova et al., Am. J. Hematol. 53 (1996) 59.

Variability  in hemoglobin F level of normal adults

C64.  John et al., Clin. Chem. 43 (1997) 968

Evaluation  of a hemoglobin analyzer, PolyCAT A as the gold standard.

C65.  Lindner et al., J. Chromatogr. A 782 (1997) 55.

Purif.  of phosphorylated histones by hydrophilic interaction  chromatography.

C66.  McDonald et al., Protein Exp. Purif. 8 (1996) 97

QC  analysis for deamidated variant of . a recombinant  fusion protein of basic fibroblast growth factor-saporin mitotoxin.

C67.  Frantzen et al., J. Chromatogr. B699 (1997) 269

 Modified  hemoglobins: A review of . analytical methods

C68.  Mario et al., Clin. Chem. 43 (1997) 2137

Comparison  of HPLC and IEF for hemoglobin variant analysis

C69.  Pearson et al., Hemoglobin 22 (1998) 245

Reversing the Hb F to Hb A switch in rats

C70.  Hazelett et al., Alcohol Clin. Exp Res.22(1998) 1813

Comparison  of hemoglobin-acetaldehyde . with CDT and other alcoholism  markers.

C71.  Sanchez et al., Thrombosis Res. 87 (1997)289.

Purif.of  chimeric fibrolase attached to an RGD-like peptide

C72.  Lindner et al., J. Cancer Res. Clin. Oncol, 125 (1999) 182

Deamidation  and N-terminal modification . of H1 histones as a function  of age

C73.  Levandoski et al., J. Neurochem. 74 (2000) 1279

Purification  of recombinant alpha- . bungarotoxin expressed in Pichia  pastoris.

C74.  Parsell et al., J. Biol. Chem. 271 (1996) 27936

Separation  of 32P-phosphorylated from native . relaxin

C75.  Sweet et al., J. Biol. Chem. 272 (1997) 916

Isolation  of phosphorylated Lys-C fragments of linker histones from  Tetrahymena.

C76.  Al-Abed et al., Proc. Natl. Acad. Sci. 96 (1999) 2385.

Separation  of rat hemoglobins with adducts of glucose, acetaldehyde, and  advanced glycation endproducts

C77.  Paleari et al., Clin. Chem 45 (1999) 21

HPLC  of Hemoglobin J Sardegna, with a post- translational  deamidation.

C78.  Campbell et al., Cytometry 35 (1999) 242.

HPLC  of Hemoglobin F to correlate results from flow cytometry of  erythrocytes.

C79.  Bohmer et al., Blood 95 (2000) 2967

Stimulation  of Hb F production by TGF-beta; Hbs F and A measured  by HPLC and flow cytometry

C80.  Roberts et al., Clin. Chem 46 (2000) 569.

Glycohemoglobin  measurements with Hb variants.

C81.  Bissé et al., J. Biol. Chem. 275 (2000) 21380.;

Detection  of a Thr to Ala replacement in Hb Saale

C82.  Bissé et al., J. Biol. Chem. 275 (2000) 6234.

Purification  of a Fe-S cluster transcription factor protein, FNR,  from E. coli.

C83.  Theodorakis et al., Biochim. Biophys. Acta 1252(1995) 14.

Cytochrome  C derivatives with thiopropyl groups at Lys-; separation of  positional variants

C84.  Bates et al., J. Biol. Chem. 275 (2000) 6234

Purification  of E. coli transcription factor FNR mutants

C85.  Xu et al., J. Biol. Chem. 275 (2000) 17086

Purification  of restriction endonuclease Hpy1881 from Helicobacter pylori.

C86.  Zhang et al., J. Chromatogr. B 759 (2001) 1

Chromatography  vs ESI-MS: effect of hemoglobin variants on measurement of glycation

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