The Experts below are selected from a list of 31095 Experts worldwide ranked by ideXlab platform
Tony G Moreno - One of the best experts on this subject based on the ideXlab platform.
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validatIon of a ph gradient based Ion Exchange Chromatography method for high resolutIon monoclonal antibody charge variant separatIons
Journal of Pharmaceutical and Biomedical Analysis, 2011Co-Authors: Tony G Moreno, Dell FarnanAbstract:Ion-Exchange Chromatography is widely used for profiling the charge heterogeneity of proteins, including monoclonal antibodies. Despite good resolving power and robustness, Ionic strength-based Ion-Exchange separatIons are product-specific and time-consuming to develop. We have previously reported a novel pH-based separatIon of proteins by catIon Exchange Chromatography that was multi-product, high-resolutIon, and robust against variatIons in sample matrix salt concentratIon and pH. In this study, a pH gradient-based separatIon method using catIon Exchange Chromatography was evaluated in a mock validatIon. This method was shown to be robust for monoclonal antibodies and suitable for its intended purpose of charge heterogeneity analysis. Simple mixtures of defined buffer components were used to generate the pH gradients that separated closely related antibody species. ValidatIon characteristics, such as precisIon and linearity, were evaluated. Robustness to changes in protein load, buffer pH and column oven temperature was demonstrated. The stability-indicating capability of this method was determined using thermally stressed antibody samples. In additIon, intermediate precisIon was demonstrated using multiple instruments, multiple analysts, multiple column lots, and different column manufacturers. Finally, the precisIon for this method was compared to conventIonal Ion-Exchange Chromatography and imaged capillary isoelectric focusing. These results demonstrate the superior precisIon and robustness of this multi-product method, which can be used for the high-throughput evaluatIon of in-process and final product samples.
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multiproduct high resolutIon monoclonal antibody charge variant separatIons by ph gradient Ion Exchange Chromatography
Analytical Chemistry, 2009Co-Authors: Dell Farnan, Tony G MorenoAbstract:In the biotechnology industry, Ion-Exchange Chromatography is widely used for profiling the charge heterogeneity of proteins, including monoclonal antibodies. Ionic strength based Ion Exchange separatIons, while having excellent resolving power and robustness, are product specific and time-consuming to develop. In the present work, a pH gradient based separatIon using a catIon Exchange column is described and shown to be a multiproduct charge sensitive separatIon method for monoclonal antibodies. Simple mixtures of defined buffer components were used to generate the pH-gradients that separate closely related antibody species. The form of the pH gradient was controlled and optimized by the pump as well as the buffer compositIon if necessary. During this work, the buffer compositIons for the separatIon were optimized in parallel for several MAbs. The data shows that the multiproduct method is optimal for all of the MAbs studied. OperatIonal aspects of the separatIon such as column chemistry, column length, ...
Dell Farnan - One of the best experts on this subject based on the ideXlab platform.
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validatIon of a ph gradient based Ion Exchange Chromatography method for high resolutIon monoclonal antibody charge variant separatIons
Journal of Pharmaceutical and Biomedical Analysis, 2011Co-Authors: Tony G Moreno, Dell FarnanAbstract:Ion-Exchange Chromatography is widely used for profiling the charge heterogeneity of proteins, including monoclonal antibodies. Despite good resolving power and robustness, Ionic strength-based Ion-Exchange separatIons are product-specific and time-consuming to develop. We have previously reported a novel pH-based separatIon of proteins by catIon Exchange Chromatography that was multi-product, high-resolutIon, and robust against variatIons in sample matrix salt concentratIon and pH. In this study, a pH gradient-based separatIon method using catIon Exchange Chromatography was evaluated in a mock validatIon. This method was shown to be robust for monoclonal antibodies and suitable for its intended purpose of charge heterogeneity analysis. Simple mixtures of defined buffer components were used to generate the pH gradients that separated closely related antibody species. ValidatIon characteristics, such as precisIon and linearity, were evaluated. Robustness to changes in protein load, buffer pH and column oven temperature was demonstrated. The stability-indicating capability of this method was determined using thermally stressed antibody samples. In additIon, intermediate precisIon was demonstrated using multiple instruments, multiple analysts, multiple column lots, and different column manufacturers. Finally, the precisIon for this method was compared to conventIonal Ion-Exchange Chromatography and imaged capillary isoelectric focusing. These results demonstrate the superior precisIon and robustness of this multi-product method, which can be used for the high-throughput evaluatIon of in-process and final product samples.
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multiproduct high resolutIon monoclonal antibody charge variant separatIons by ph gradient Ion Exchange Chromatography
Analytical Chemistry, 2009Co-Authors: Dell Farnan, Tony G MorenoAbstract:In the biotechnology industry, Ion-Exchange Chromatography is widely used for profiling the charge heterogeneity of proteins, including monoclonal antibodies. Ionic strength based Ion Exchange separatIons, while having excellent resolving power and robustness, are product specific and time-consuming to develop. In the present work, a pH gradient based separatIon using a catIon Exchange column is described and shown to be a multiproduct charge sensitive separatIon method for monoclonal antibodies. Simple mixtures of defined buffer components were used to generate the pH-gradients that separate closely related antibody species. The form of the pH gradient was controlled and optimized by the pump as well as the buffer compositIon if necessary. During this work, the buffer compositIons for the separatIon were optimized in parallel for several MAbs. The data shows that the multiproduct method is optimal for all of the MAbs studied. OperatIonal aspects of the separatIon such as column chemistry, column length, ...
Xueguo Chen - One of the best experts on this subject based on the ideXlab platform.
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separatIon and detectIon of compounds in honeysuckle by integratIon of Ion Exchange Chromatography fractIonatIon with reversed phase liquid Chromatography atmospheric pressure chemical IonizatIon mass spectrometer and matrix assisted laser desorptIon IonizatIon time of flight mass spectrometry analysis
Journal of Pharmaceutical and Biomedical Analysis, 2006Co-Authors: Xueguo Chen, Liang Kong, Hanfa ZouAbstract:A hyphenated method for the isolatIon and identificatIon of components in a traditIonal Chinese medicine of Honeysuckle was developed. Ion-Exchange Chromatography (IEC) was chosen for the fractIonatIon of Honeysuckle extract, and then followed by concentratIon of all the fractIons with rotary vacuum evaporator. Each of the enriched fractIons was then further analyzed by reversed-phase liquid Chromatography-atmospheric pressure chemical IonizatIon mass spectrometer (RPLC-APCI/MS) and matrix-assisted laser desorptIon/IonizatIon time-of-flight mass spectrometry (MALDI-TOF/MS) with matrix of oxidized carbon nanotubes, respectively. It can be noted totally more than 117 components were detected by UV detector, APCI/MS and MALDI-TOF/MS in Honeysuckle extract except the 145 components identified by MALDI-TOF/MS alone with this integrated approach, and 7 of them were preliminary identified according to their UV spectra and mass spectra performed by APCI/MS and MALDI-TOF/MS, respectively. The obtained analytical results not only indicated the approach of integratIon IEC fractIonatIon with RPLC-APCI/MS and MALDI-TOF/MS is capable of analyzing complex samples, but also exhibited the potential power of the mass spectrometer in detectIon of low-mass compounds, such as traditIonal Chinese medicines (TCMs) and complex biological samples.
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integratIon of Ion Exchange Chromatography fractIonatIon with reversed phase liquid Chromatography atmospheric pressure chemical IonizatIon mass spectrometer and matrix assisted laser desorptIon IonizatIon time of flight mass spectrometry for isolatIon and identificatIon of compounds in psoralea corylifolia
Journal of Chromatography A, 2005Co-Authors: Xueguo Chen, Liang Kong, Xingye Su, Mingliang YeAbstract:An approach for the separatIon and identificatIon of components in a traditIonal Chinese medicine Psoralea corylifolia was developed. Ion-Exchange Chromatography (IEC) was applied for the fractIonatIon of P. corylifolia extract, and then followed by concentratIon of all the fractIons with rotary vacuum evaporator. Each of the enriched fractIons was then further separated on an ODS column with detectIon of UV absorbance and atmospheric pressure chemical IonizatIon mass spectrometer (APCI/MS), respectively, and also analyzed by matrix-assisted laser desorptIon/IonizatIon time-of-flight mass spectrometry (MALDI-TOF/MS) with matrix of oxidized carbon nanotubes. Totally more than 188 components in P. corylifolia extract were detected with this integrated approach, and 12 of them were preliminary identified according to their UV spectra and mass spectra performed by APCI/MS and MALDI-TOF/MS. The obtained analytical results not only demonstrated the powerful resolutIon of integratIon IEC fractIonatIon with reversed-phase liquid Chromatography (RPLC)-APCI/MS and MALDI-TOF/MS for analysis of compounds in a complex sample, but also exhibited the superiority of APCI/MS and MALDI-TOF/MS for identificatIon of low-mass compounds, such as for study of traditIonal Chinese medicines (TCMs) and metabolome.
Hanfa Zou - One of the best experts on this subject based on the ideXlab platform.
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separatIon and detectIon of compounds in honeysuckle by integratIon of Ion Exchange Chromatography fractIonatIon with reversed phase liquid Chromatography atmospheric pressure chemical IonizatIon mass spectrometer and matrix assisted laser desorptIon IonizatIon time of flight mass spectrometry analysis
Journal of Pharmaceutical and Biomedical Analysis, 2006Co-Authors: Xueguo Chen, Liang Kong, Hanfa ZouAbstract:A hyphenated method for the isolatIon and identificatIon of components in a traditIonal Chinese medicine of Honeysuckle was developed. Ion-Exchange Chromatography (IEC) was chosen for the fractIonatIon of Honeysuckle extract, and then followed by concentratIon of all the fractIons with rotary vacuum evaporator. Each of the enriched fractIons was then further analyzed by reversed-phase liquid Chromatography-atmospheric pressure chemical IonizatIon mass spectrometer (RPLC-APCI/MS) and matrix-assisted laser desorptIon/IonizatIon time-of-flight mass spectrometry (MALDI-TOF/MS) with matrix of oxidized carbon nanotubes, respectively. It can be noted totally more than 117 components were detected by UV detector, APCI/MS and MALDI-TOF/MS in Honeysuckle extract except the 145 components identified by MALDI-TOF/MS alone with this integrated approach, and 7 of them were preliminary identified according to their UV spectra and mass spectra performed by APCI/MS and MALDI-TOF/MS, respectively. The obtained analytical results not only indicated the approach of integratIon IEC fractIonatIon with RPLC-APCI/MS and MALDI-TOF/MS is capable of analyzing complex samples, but also exhibited the potential power of the mass spectrometer in detectIon of low-mass compounds, such as traditIonal Chinese medicines (TCMs) and complex biological samples.
G. Chevreux - One of the best experts on this subject based on the ideXlab platform.
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Charge variants characterizatIon of a monoclonal antibody by Ion Exchange Chromatography coupled on-line to native mass spectrometry: Case study after a long-term storage at +5 °C
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 2017Co-Authors: Y. Leblanc, N. Bihoreau, Cesar Ramon, G. ChevreuxAbstract:Numerous putative post-translatIonal modificatIons may induce variatIons of monoclonal antibodies charge distributIon that can potentially affect their biological activity. The characterizatIon and the monitoring of these charge variants are critical quality requirements to ensure stability and process consistency. Charge variants are usually characterized by preparative Ion Exchange Chromatography, collectIon of fractIons and subsequent reverse-phase liquid Chromatography with mass spectrometry analysis. While this process can be automatized by on-line two-dimensIonal Chromatography, it remains often complex and time consuming. For this reason, a straightforward on-line charge variant analysis method is highly desirable and analytical laboratories are actively pursuing efforts to overcome this challenge. In this study, a mixed mode Ion Exchange chromatographic method using volatile salts and coupled on-line to native mass spectrometry was developed in associatIon with a middle-up approach for a detailed characterizatIon of monoclonal antibodies charge variants. An aged monoclonal antibody, presenting a complex charge variant profile was successfully investigated by this methodology as a case study. Results demonstrate that deamidatIon of the heavy chain was the major degradatIon pathway after long-term storage at 5 °C while oxidatIon was rather low. The method was also very useful to identify all the clipped forms of the antibody.
