2019
DOI: 10.1007/s00216-019-02029-0
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A rapid and quantitative technique for assessing IgG monomeric purity, calibrated with the NISTmAb reference material

Abstract: The fraction of intact monomer in a sample (moles/moles), the monomeric purity, is measured as a quality control in therapeutic monoclonal antibodies but is often unknown in research samples and remains a major source of variation in quantitative antibody-based techniques such as immunoassay development. Here, we describe a novel multiplex technique for estimating the monomeric purity and antigen affinity of research grade antibody samples. Light scattering was used to simultaneously observe the mass of antibo… Show more

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Cited by 6 publications
(8 citation statements)
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“…The hydrodynamic size of the native IgG was found to be ∼5 nm, in agreement with the literature ( Hawe et al, 2011 ). The absorbance of native IgG was 0.172, which corresponded to 0.1 mg/mL of IgG using Beer Lambert's law ( Reader et al, 2019 ). The effects of varying temperature and combined UV and temperature with time were evaluated at the conditions suggested by Design-Expert software ( Table 2 ).…”
Section: Resultsmentioning
confidence: 99%
See 1 more Smart Citation
“…The hydrodynamic size of the native IgG was found to be ∼5 nm, in agreement with the literature ( Hawe et al, 2011 ). The absorbance of native IgG was 0.172, which corresponded to 0.1 mg/mL of IgG using Beer Lambert's law ( Reader et al, 2019 ). The effects of varying temperature and combined UV and temperature with time were evaluated at the conditions suggested by Design-Expert software ( Table 2 ).…”
Section: Resultsmentioning
confidence: 99%
“…To estimate the final concentration of the stock solution, the absorbance values were measured using a UV spectrophotometer (Thermo Scientific; Evolution 201) at 280 nm. The stock concentration was calculated using Beer Lambert's law with the extinction coefficient value of 210,000 M -1 cm -1 ( Reader et al, 2019 ). A final concentration of 0.1 mg/mL was prepared as a working solution for further usage.…”
Section: Methodsmentioning
confidence: 99%
“…However, the antibody orientation on the sensor surface can be interrogated using an Fc-presenting assay by passing PAG over the surface. The surface density and orientation of antibodies produce a measure of quality control in the manufacture of the assay arrays 19 .…”
Section: Experimental Methods and Materialsmentioning
confidence: 99%
“…The assay platform has been optimised for two assays, C-reactive protein (CRP) and total IgG both have commercial interest. Total IgG provides useful information about antibody integrity 19 and in-line QC of the materials on delivery supporting both the CRP assay and the total IgG assay. CRP is the target clinical assay and is the subject of ~5,000 publications per year (PubMed) as a sensitive but not specific biomarker of systemic inflammation in many conditions such as sepsis 20 .…”
Section: Introductionmentioning
confidence: 99%
“…The tests were performed on the portable bench-top multiplexed array technology that has been described in detail elsewhere. [6][7][8][9][10] It has been shown effective at detecting antibody in response to vaccination 11 and has characterised accuracy and precision for CRP and total IgG assays 6 with typically 10% accuracy and intra-day precision of less than 5%. The technology consists of an array of 170 of gold nanoparticle spots which scatter light into a video camera when illuminated from below ( Fig.…”
Section: Multiplexed Covid-19 Antigen Array and Liscar Readermentioning
confidence: 99%