High-resolution chromatography of nucleic acids on the gen-pak fax column

Stowers, Deborah J.; Keim, Jenny M.B.; Paul, P. S.; Lyoo, Young S.; Merion, Michael; Benbow, Robert M.

doi:10.1016/s0021-9673(01)94008-7

Cited by 28 publications

(7 citation statements)

References 25 publications

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“…In a previous report from our laboratory (17), we presented preliminary evidence indicating that the tyrosine at position 37 NSDSE NSYPG NSNTG QDAPG VVSHFND VVSHFNK NRKKKNP DIPAIRL IIKRIKL IVLHVKV vC vC vC vC TC TC EC DC TC TC 30 MYI (4) or by ion exchange on high-performance liquid chromatography (HPLC), using a Waters Gen-Pak FAX column (38).…”

mentioning

confidence: 99%

Aromaticity at position 37 in human epidermal growth factor is not obligatory for activity.

Engler¹,

Hauser²,

Cook³

et al. 1991

Mol. Cell. Biol.

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The third disulfide loop (amino acids 33 to 42) of human epidermal growth factor (hEGF) encompasses the region of highest amino acid conservation among all of the EGF-like family of molecules. The importance of some of these highly conserved residues for the maintenance of biological activity, especially the aromatic amino acid tyrosine at position 37, has until now been considered essential on the basis of previous studies with the EGF-like molecule transforming growth factor alpha. Variants at the Tyr-37 position of hEGF were constructed by site-directed mutagenesis. The substituting amino acids were phenylalanine, histidine, serine, alanine, aspartic acid, arginine, and glycine. The variants were tested for their ability to competitively displace native ['25I]hEGF from its receptor and to stimulate the protein-tyrosine kinase activity of the receptor; the order of efficacy of substituting amino acids was Phe > His > Ser > Ala > Asp > Arg > Gly in both assays. All were effective, with no or only moderate reduction in potency, in stimulating the incorporation of [3H]thymidine into acid-insoluble material of quiescent mouse A31 cells. Only Tyr-37-*Ala, Tyr-37--Arg and Tyr-37-*Gly were slightly less potent in the cell assay. Thus, neither tyrosine nor another aromatic amino acid at position 37 in hEGF is essential for full biological activity.

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mentioning

confidence: 99%

Aromaticity at position 37 in human epidermal growth factor is not obligatory for activity.

Engler¹,

Hauser²,

Cook³

et al. 1991

Mol. Cell. Biol.

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“…For practical and economical reasons it is rarely attractive to use oligoprobes requiring more than 50-70 coupling cycles. Full-length sequences can be purified from failure sequences and damaged DNA using reverse-phase (RP) and/or ion-exchange HPLC (Stowers et al 1988;Larsson andHougaard 1990, 1993). Synthetic probes may be chemically or enzymatically tailed with reporter molecules.…”

Section: Production Of Probesmentioning

confidence: 99%

Non-radioactive in situ hybridization for mRNA with emphasis on the use of oligodeoxynucleotide probes

Hougaard

Hansen

Larsson

1997

Histochemistry and Cell Biology

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“…Separation of the different forms of DNA is possible using ion-exchange chromatography [8] and a method was established to characterise the pDNA using ionexchange HPLC with a non-porous anion exchange resin (TSK Gel DEAE-NPR) however the presence of the lipid components interfered with the separation when analysing Complex. The use of capillary electrophoresis with an entangled polymer matrix to analyse DNA is well documented using a variety of polymeric media to effect the size based separation [9][10][11][12][13][14][15][16][17][18] and much research is being invested to elucidate the mechanisms involved in the separations obtained [ 19,20].…”

Section: Introductionmentioning

confidence: 99%

The use of capillary electrophoresis with entangled polymer matrix to analyse plasmid DNA and a cationic lipid/cholesterol liposome: DNA complex

1999

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SummaryA liposome based gene therapeutic product is being developed for the treatment of cystic fibrosis. The product comprises a complex of plasmid DNA and a cationic lipid/cholesterol liposome. Using CE with an entangled polymer matrix, routine separations of linear, supercoiled and open-circle conformers of DNA in plasmid DNA and the liposome complex have been performed. The CE method has been used to support novel quality control and process validation for the manufacture of plasmid DNA and to monitor the degradation of DNA in the liposome complex. Significant features of the method are simple sample preparation and the use of direct UV detection, avoiding the use of potentially mutagenic reagents.

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High-resolution chromatography of nucleic acids on the gen-pak fax column

Cited by 28 publications

References 25 publications

Aromaticity at position 37 in human epidermal growth factor is not obligatory for activity.

Aromaticity at position 37 in human epidermal growth factor is not obligatory for activity.

Non-radioactive in situ hybridization for mRNA with emphasis on the use of oligodeoxynucleotide probes

The use of capillary electrophoresis with entangled polymer matrix to analyse plasmid DNA and a cationic lipid/cholesterol liposome: DNA complex

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