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Titolo:
A method for routine analysis of recombinant immunoglobulins (rIgGs) by capillary isoelectric focusing (cIEF)
Autore:
Tang, S; Nesta, DP; Maneri, LR; Anumula, KR;
Indirizzi:
SmithKlineia,echam Pharmaceut, Res & Dev, Dept Bioanalyt Sci, King Of Pruss SmithKline Beecham Pharmaceut King Of Prussia PA USA 19406 King Of Pruss SmithKlinePrussia, Pharmaceut, Res & Dev, Dept Pharmaceut Technol, King OfSmithKline Beecham Pharmaceut King Of Prussia PA USA 19406 chnol, King Of
Titolo Testata:
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
fascicolo: 3-4, volume: 19, anno: 1999,
pagine: 569 - 583
SICI:
0731-7085(199903)19:3-4<569:AMFRAO>2.0.ZU;2-K
Fonte:
ISI
Lingua:
ENG
Soggetto:
PROTEINS; ELECTROPHORESIS;
Keywords:
recombinant IgG; isoelectric focusing; capillary electrophoresis;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
10
Recensione:
Indirizzi per estratti:
Indirizzo: Anumula, KR SmithKlineia,echam Pharmaceut, Res & Dev, Dept Bioanalyt Sci, King Of Pruss SmithKline Beecham Pharmaceut King Of Prussia PA USA 19406 ss
Citazione:
S. Tang et al., "A method for routine analysis of recombinant immunoglobulins (rIgGs) by capillary isoelectric focusing (cIEF)", J PHARM B, 19(3-4), 1999, pp. 569-583

Abstract

A capillary isoelectric focusing (cIEF) method was developed for routine analysis of recombinant immunoglobulins (rIgGs). The cIEF method used a dimethyl siloxane-coated capillary and a separation matrix of 2% ampholytes in 0.4% methylcellulose (MC). The rIgGs, and internal pi marker protein standards, were mixed with carrier ampholyte in MC, focused using high voltage, and then the protein bands were mobilized past a UV detector by simultaneousapplication of low pressure and voltage. Qualitatively and quantitatively equivalent rIgG focusing profiles were obtained via cIEF and gel-based IEF,with individual isoform peak area percentages and calculated peak pI values being comparable for the same samples. Linear relationships were obtainedfor peak area response versus sample concentration, and for the pI gradient developed between the internal pr marker standards. The relative standarddeviation (RSD) in rIgG peak areas was less than 2% intra-day and less than 8% inter-day (72 h). The RSD for the mobilization times of rIgG peaks wasless than 1% intra-day and less than 3% inter-day (72 h). There was no observed decrease in the performance of the capillary over 150 analyses. cIEF offers several important advantages over gel IEF, e.g. direct, quantitativedetection of proteins by intrinsic UV absorbance at 280 nm, rapid analyses(less than or equal to 30 min), capability of automation, and one-step, electronic data analysis and archival. These data demonstrate the superiorityof the cIEF method for routine analysis of rIgGs. (C) 1999 Elsevier Science B.V. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 05/04/20 alle ore 03:41:05