Purification of antibody Fab fragments by cation-exchange chromatography and pH gradient elution

Mhatre, Rohin; Nashabeh, Wassim; Schmalzing, Dieter; Yao, Xian-Wei; Fuchs, Martin; Whitney, Duncan; Regnier, Fred E.

doi:10.1016/0021-9673(95)00319-i

Cited by 34 publications

(13 citation statements)

References 15 publications

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“…Al1 three Fab fragments displayed equal immunogenic activity to cortisol, ieading to the formation of three complex peaks. To simplify the electrophoretic pattern, the three Fab fragments were fractionated based on their p l differences by HPLC on a cation exchanger [27]. A single Fab fragment was then used in the assay.…”

Section: Introductionmentioning

confidence: 99%

Capillary electrophoresis based immunoassays: A critical review

Schmalzing

Nashabeh²

1997

Electrophoresis

102

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The purpose of this article is to review and evaluate the uses and potentials of capillary electrophoresis (CE) in immunoassay analysis (IA). This review will be divided into four sections. First, a brief summary of the fundamentals, applications and requirements of immunoassays in both research and clinical diagnostics will be given. This section will also cover the rationale behind the current research to combine CE and IA. Also, the specific needs to CE in this field will be addressed. Second, the modes of use of CE in IA will be discussed and typical applications for each will be given. Third, a separate section will be devoted to the investigation of performing immunoassays on micro fabricated devices, an interesting alternative to the conventional capillary-based approach. Finally, a critical assessment of the current status and merits of this technology will be presented.

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Section: Introductionmentioning

confidence: 99%

Capillary electrophoresis based immunoassays: A critical review

Schmalzing

Nashabeh²

1997

Electrophoresis

102

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“…Generally, a linear salt or pH gradient elution program can be conducted to determine the best elution condition. [55][56][57][58] An elution condition development study conducted for an IgG1 is shown in Figure 6. Four runs were performed in this study with two Fractogel cation exchange resins, Fractogel COO -(weak cation exchanger) chromatographic step such as cation exchange, so it may not be applicable to all products.…”

Section: Chromatographic Processesmentioning

confidence: 99%

Recovery and purification process development for monoclonal antibody production

Liu

Winter

et al. 2010

mAbs

463

411

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“…Variation in the extent of C-terminal processing can lead to production lots with different charge distributions (5). The charge variants, which are generated by incomplete posttranslational cleavage of the C-terminal lysines, can be detected by CIEX or cIEF, although additional factors, such as deamidation, phosphorylation, or sialic acid variability, may also contribute to the overall charge heterogeneity (1)(2)(3)(4)(5).…”

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confidence: 99%

“…D2E7 is a 148-kDa IgG containing two heavy chains (50.6 kDa each) and two light chains (23.4 kDa each). It is secreted from recombinant Chinese hamster ovary (CHO) 2 cells grown in a stirred tank reactor, recovered from the supernatant by micro-and ultrafiltration, and purified to homogeneity by column chromatography. Variation in the extent of C-terminal processing can lead to production lots with different charge distributions (5).…”

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confidence: 99%