Separation of recombinant human growth hormone from Escherichia coli cell pellet extract by capillary zone electrophoresis

McNerney, Thomas; Watson, S. K.; Sim, Jek-Hui; Bridenbaugh, Robert

doi:10.1016/0021-9673(96)00421-9

Cited by 39 publications

(9 citation statements)

References 10 publications

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“…CE analyses of VIP were first carried out using a fused-silica capillary with various buffers at pH 7.2 (Tris-HCl, Tricine, phosphate). Whatever the buffer used, no peak was observed under these conditions even with phosphate buffer, which is believed to reduce peptide adsorption [53,54]. Consequently, to overcome the drawbacks associated with untreated fused-silica capillaries, neutral-coated capillaries (PAA and PEO) and positively charged ones (PDDA) were investigated.…”

Section: Use Of Different Capillaries For the Analysis Of Vip By Cementioning

confidence: 99%

Determination of binding constants of vasoactive intestinal peptide to poly(amidoamine) dendrimers designed for drug delivery using ACE

et al. 2007

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The purpose of the present paper was to study at physiological pH the affinity between vasoactive intestinal peptide (VIP) and four poly(amidoamine) dendrimers (PAMAMs) designed for drug delivery. Therefore, a fast and reproducible CE method was first developed to analyze the strongly basic peptide. To allow an accurate determination of binding constant (K) values, the ability to suppress peptide adsorption onto the silica capillary of nonpermanent coatings (poly(ethylene oxide) (PEO), low and medium relative molecular masses poly(diallyldimethylammonium chloride) (PDDA)) or poly(acrylamide) permanent coating (PAA) was evaluated. Very good intraday repeatability of VIP migration times and peak areas (0.1-0.6 and 2.9-4.9% RSD, respectively) was obtained using two of the investigated coatings (PEO and PDDA with medium molecular mass). ACE combined with these dynamic coatings was then employed to evaluate K between VIP and two amine-terminated PAMAM dendrimers of generation 2 and 5 (G2.NH2, G5.NH2) and two carboxyl-terminated PAMAM derivatives of generation 2 and 5 (G2.COOH, G5.COOH). Binding constant of (6.7 +/- 1.1) x 10(4)/M could be determined for the couple VIP/G5.NH2, while no affinity was evidenced between VIP and all other dendrimers investigated. These results suggest that G5.NH2 might be an interesting carrier for the delivery of VIP.

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Section: Use Of Different Capillaries For the Analysis Of Vip By Cementioning

confidence: 99%

Determination of binding constants of vasoactive intestinal peptide to poly(amidoamine) dendrimers designed for drug delivery using ACE

et al. 2007

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“…However, kin17 is a basic protein and exhibited a strong interaction with the silica capillary wall at neutral pH (data not shown). To minimize proteincapillary surface interactions, CE analyses of kin17 protein were first carried out using a fused-silica capillary with phosphate buffer as the beneficial effect of phosphate to reduce protein adsorption has been discussed several times [29][30][31]. But no peak was observed in these conditions.…”

Section: Analysis With a Fused-silica Capillarymentioning

confidence: 99%

Poly(ethylene oxide) facilitates the characterization of an affinity between strongly basic proteins with DNA by affinity capillary electrophoresis

et al. 2005

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In order to study kin17 protein-DNA affinity, we have developed a fast and reproducible capillary electrophoresis (CE) analysis of a strongly basic protein: kin17 protein, using a nonpermanent coating based on poly(ethylene oxide) (PEO) to avoid adsorption of kin17. The coating procedure was optimized to provide a residual and stable electroosmotic flow (EOF = 5 x 10(-5) cm(2)/V x s), exhibiting RSD of 0.3% and excellent long-term stability. Good intraday and interday reproducibility of kin17 migration times (0.8 and 0.3% relative standard deviation (RSD), respectively) enabled us to consider that the recovery percentage obtained for kin17 protein was satisfactory (79%). The potential of this PEO-based coating procedure was evaluated for affinity CE method in order to study the affinity of kin17 protein for two single-stranded DNA (ssDNA) models: polydeoxyadenylic acid and polydeoxycytidilic acid (pdA and pdC). Binding constants (1.5 x 10(7) +/- 17% and 1.7 x 10(7) + 25%M(-1)) were evaluated assuming a 1:1 affinity between kin17 and pdA or pdC, respectively.

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“…Such examples will be mostly limited to proteins of biomedical and biotechnological interest. McNerney et al [87] reported the analysis of recombinant human growth hormone (rHGH) in E. coli cell pellets by the use of phosphate-deactivated capillaries. They could resolve a number of variants, such as Des-Phe, des Phe-Pro, mono-deamidated and di-deamidated HGH forms.…”

Section: Ce Analysis Of Proteinsmentioning

confidence: 99%

Capillary electrophoretic analysis of proteins and peptides of biomedical and pharmacological interest

Righetti

2001

Biopharm & Drug Disp

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Capillary electrophoresis (CE) is an automated approach to electrokinetic separations that has had a deep impact in all fields of life sciences, including biomedical and biotechnological research and clinical and forensic practice. The present review highlights aspects of peptides and proteins separations, with particular emphasis on macromolecular analytes of biomedical interest. Among the various CE techniques available, a novel methodology is here illustrated consisting in separations in acidic, isoelectric buffers, which have the advantage of protonating the silica wall, thus minimizing interactions of proteinaceous material with the siliceous surface, while allowing delivery of high voltage gradients, due to their low conductivities. The review ends with applications of CE to the analysis of folding/unfolding/refolding/misfolding of proteins, a field which has deep implications in the biomedical arena, since it is connected to a host of disorders, such as prion protein diseases.

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Separation of recombinant human growth hormone from Escherichia coli cell pellet extract by capillary zone electrophoresis

Cited by 39 publications

References 10 publications

Determination of binding constants of vasoactive intestinal peptide to poly(amidoamine) dendrimers designed for drug delivery using ACE

Determination of binding constants of vasoactive intestinal peptide to poly(amidoamine) dendrimers designed for drug delivery using ACE

Poly(ethylene oxide) facilitates the characterization of an affinity between strongly basic proteins with DNA by affinity capillary electrophoresis

Capillary electrophoretic analysis of proteins and peptides of biomedical and pharmacological interest

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