Development of a validated liquid chromatography tandem mass spectrometry assay for a PEGylated adnectin in cynomolgus monkey plasma using protein precipitation and trypsin digestion

Dawes, Michelle L.; Gu, Huidong; Wang, Jian; Schuster, Alan; Haulenbeek, Jonathan

doi:10.1016/j.jchromb.2013.06.027

Cited by 20 publications

(10 citation statements)

References 27 publications

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“…A full method validation consists of at least three accuracy and precision runs, which can provide sufficient instrument response data from STD or QC samples for the calculation of σ. Figure b and Figure a–c ,, show the relationships between σ and x (blue line), and between σ 2 and x (red line) from STD curve instrument responses from four assay validations. A quick eyeball of these plots suggested that 1/ x 2 should be used as the weighting factor for 3 out of 4 assays listed, as very good straight line relationships between σ and x and concave upward quadratic relationships between σ 2 and x were found over their corresponding STD ranges.…”

Section: Results and Discussionmentioning

confidence: 99%

Selecting the Correct Weighting Factors for Linear and Quadratic Calibration Curves with Least-Squares Regression Algorithm in Bioanalytical LC-MS/MS Assays and Impacts of Using Incorrect Weighting Factors on Curve Stability, Data Quality, and Assay Performance

et al. 2014

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A simple procedure for selecting the correct weighting factors for linear and quadratic calibration curves with least-squares regression algorithm in bioanalytical LC-MS/MS assays is reported. The correct weighting factor is determined by the relationship between the standard deviation of instrument responses (σ) and the concentrations (x). The weighting factor of 1, 1/x, or 1/x(2) should be selected if, over the entire concentration range, σ is a constant, σ(2) is proportional to x, or σ is proportional to x, respectively. For the first time, we demonstrated with detailed scientific reasoning, solid historical data, and convincing justification that 1/x(2) should always be used as the weighting factor for all bioanalytical LC-MS/MS assays. The impacts of using incorrect weighting factors on curve stability, data quality, and assay performance were thoroughly investigated. It was found that the most stable curve could be obtained when the correct weighting factor was used, whereas other curves using incorrect weighting factors were unstable. It was also found that there was a very insignificant impact on the concentrations reported with calibration curves using incorrect weighting factors as the concentrations were always reported with the passing curves which actually overlapped with or were very close to the curves using the correct weighting factor. However, the use of incorrect weighting factors did impact the assay performance significantly. Finally, the difference between the weighting factors of 1/x(2) and 1/y(2) was discussed. All of the findings can be generalized and applied into other quantitative analysis techniques using calibration curves with weighted least-squares regression algorithm.

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Section: Results and Discussionmentioning

confidence: 99%

Selecting the Correct Weighting Factors for Linear and Quadratic Calibration Curves with Least-Squares Regression Algorithm in Bioanalytical LC-MS/MS Assays and Impacts of Using Incorrect Weighting Factors on Curve Stability, Data Quality, and Assay Performance

et al. 2014

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“…LC–MRM also has been applied to the quantification of PEGylated proteins [198,201,202]. Usually, the MW of intact PEGylated protein is beyond the range of detection.…”

Section: Bioanalytical Methods For Polyethylene Glycols and Pegylatedmentioning

confidence: 99%

Recent advances in the bioanalytical methods of polyethylene glycols and PEGylated pharmaceuticals

Zhang

Song

et al. 2020

J of Separation Science

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Polyethylene glycols are synthetic polymers composed of repeating oxyethylene subunits, which have been known for non-toxic, non-immunogenic, non-antigenic, good solubility in water and therefore approved for pharmaceutical applications. Recently, attachment or amalgamation of polyethylene glycols to therapeutic small molecules, peptides, proteins, or nanoparticles has become a mature technology for the sake of improving their pharmacokinetic and pharmacological profiles, also referred to as PEGylation. By comparison, there are only a few PEGylated pharmaceuticals have been registered for further clinical trials and even less was approved for marketing. High failure rate of PEGylated pharmaceuticals in pre-clinical and clinical trials could be majorly attributed to their unclear pharmacokinetic behaviors. Therefore, the in vivo fate of the PEGylated pharmaceuticals for the various routes of administration needs to be thoroughly investigated An accurate in vivo pharmacological study thereof highly depends on the precise detection of polyethylene glycols as well as their fragments in biological matrixes. The goal of this review is to highlight the analytical methods that were developed and applied to evaluate the polyethylene glycols in pharmaceutical ingredients and excipients, which bring us closer to bridging

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“…Of all classes of protein therapeutics, PEGylated proteins with high PEG content are particularly well suited to LC-MS/MS analysis because they can be extracted fairly selectively from serum via a simple protein precipitation procedure [13,16,17]. In this case, the 40 kDa PEG constitutes 77% of total molecular weight and makes the PEGylated dAb soluble in organic solvent.…”

Section: Protein Precipitation With Acid Dissociation Pretreatmentmentioning

confidence: 99%

“…In addition, the development and validation of an LC-MS/MS assay can usually be achieved in a relatively short period of time since there is no need for specific antibody reagents. Several LC-MS/MS assays have been recently developed and validated for therapeutic proteins [11][12][13][14] and the data generated by the LC-MS/MS assays were comparable with that of the LBAs. However, the data generated by both assays is not expected to be comparable in cases where the LBA format measures free drug and the LC-MS/MS measures total; or in situations where certain levels of antidrug antibodies (ADAs), soluble target or other matrix components show interference in only one of the formats.…”

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confidence: 94%

“…A new bioanalytical assay with a lower LLOQ for better characterization of PK parameters was required to support the multiple ascending dose (MAD) study. PEGylated proteins have previously been reported to be easily extracted from serum using protein precipitation with organic solvents and are suitable for LC-MS/MS analysis [13,[16][17]. LLOQs of 10-50 ng/ml were reported in all cases.…”

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confidence: 95%

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Development and Validation of an LC–MS/MS Assay for The Quantitation of A Pegylated Anti-Cd28 Domain Antibody in Human Serum: Overcoming Interference from Antidrug Antibodies and Soluble Target

et al. 2014

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Development of a validated liquid chromatography tandem mass spectrometry assay for a PEGylated adnectin in cynomolgus monkey plasma using protein precipitation and trypsin digestion

Cited by 20 publications

References 27 publications

Selecting the Correct Weighting Factors for Linear and Quadratic Calibration Curves with Least-Squares Regression Algorithm in Bioanalytical LC-MS/MS Assays and Impacts of Using Incorrect Weighting Factors on Curve Stability, Data Quality, and Assay Performance

Selecting the Correct Weighting Factors for Linear and Quadratic Calibration Curves with Least-Squares Regression Algorithm in Bioanalytical LC-MS/MS Assays and Impacts of Using Incorrect Weighting Factors on Curve Stability, Data Quality, and Assay Performance

Recent advances in the bioanalytical methods of polyethylene glycols and PEGylated pharmaceuticals

Development and Validation of an LC–MS/MS Assay for The Quantitation of A Pegylated Anti-Cd28 Domain Antibody in Human Serum: Overcoming Interference from Antidrug Antibodies and Soluble Target

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