Evaluation of Two Novel Scale-Down Devices for Testing Monoclonal Antibody Aggregation During Large-Scale Freezing

Abstract Purpose Scale-down devices (SDD) are designed to simulate large-scale thawing of protein drug substance, but require only a fraction of the material. To evaluate the performance of a new SDD that aims to predict thawing in large-scale 2 L bottles, we characterised 3D temperature profiles and changes in concentration and density in comparison to 125 mL and 2 L bottles. Differences in diffusion between a monoclonal antibody (mAb) and histidine buffer after thawing were examined. Methods Temperature profiles at six distinct positions were recorded with type T thermocouples. Size-exclusion chromatography allowed quantification of mAb and histidine. Polysorbate 80 was quantified using a fluorescent dye assay. In addition, the solution’s density at different locations in bottles and the SDD was identified. Results The temperature profiles in the SDD and the large-scale 2 L bottle during thawing were similar. Significant concentration gradients were detected in the 2 L bottle leading to marked density gradients. The SDD slightly overestimated the dilution in the top region and the maximum concentrations at the bottom. Fast diffusion resulted in rapid equilibration of histidine. Conclusion The innovative SDD allows a realistic characterisation and helps to understand thawing processes of mAb solutions in large-scale 2 L bottles. Only a fraction of material is needed to gain insights into the thawing behaviour that is associated with several possible detrimental limitations.

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Development and evaluation of a monoclonal antibody-based competitive ELISA for the detection of antibodies against H7 avian influenza virus

BMC Veterinary Research ◽

10.1186/s12917-021-02772-6 ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Yuan Li ◽

Hongliu Ye ◽

Meng Liu ◽

Suquan Song ◽

Jin Chen ◽

...

Keyword(s):

Monoclonal Antibody ◽

Influenza Virus ◽

Avian Influenza ◽

Avian Influenza Virus ◽

Large Scale ◽

Operation Time ◽

Reference Method ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Antibody Concentration

Abstract Background H7 subtype avian influenza has caused great concern in the global poultry industry and public health. The conventional serological subtype-specific diagnostics is implemented by hemagglutination inhibition (HI) assay despite lengthy operation time. In this study, an efficient, rapid and high-throughput competitive enzyme-linked immunosorbent assay (cELISA) was developed for detection of antibodies against H7 avian influenza virus (AIV) based on a novel monoclonal antibody specific to the hemagglutinin (HA) protein of H7 AIV. Results The reaction parameters including antigen coating concentration, monoclonal antibody concentration and serum dilution ratio were optimized for H7 antibody detection. The specificity of the cELISA was tested using antisera against H1 ~ H9, H11 ~ H14 AIVs and other avian viruses. The selected cut-off values of inhibition rates for chicken, duck and peacock sera were 30.11, 26.85 and 45.66% by receiver-operating characteristic (ROC) curve analysis, respectively. With HI test as the reference method, the minimum detection limits for chicken, duck and peacock positive serum reached 20, 21 and 2− 1 HI titer, respectively. Compared to HI test, the diagnostic accuracy reached 100, 98.6, and 99.3% for chicken, duck and peacock by testing a total of 400 clinical serum samples, respectively. Conclusions In summary, the cELISA assay developed in this study provided a reliable, specific, sensitive and species-independent serological technique for rapid detection of H7 antibody, which was applicable for large-scale serological surveillance and vaccination efficacy evaluation programs.

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Application of projection-based model reduction to finite-element plate models for two-dimensional traveling waves

Journal of Intelligent Material Systems and Structures ◽

10.1177/1045389x16679295 ◽

2016 ◽

Vol 28 (14) ◽

pp. 1886-1904 ◽

Cited By ~ 6

Author(s):

Vijaya VN Sriram Malladi ◽

Mohammad I Albakri ◽

Serkan Gugercin ◽

Pablo A Tarazaga

Keyword(s):

Finite Element ◽

Model Reduction ◽

Traveling Waves ◽

Large Scale ◽

Fe Model ◽

Plate Model ◽

Reduction Techniques ◽

State Frequency ◽

Scale Down ◽

The Stability

A finite element (FE) model simulates an unconstrained aluminum thin plate to which four macro-fiber composites are bonded. This plate model is experimentally validated for single and multiple inputs. While a single input excitation results in the frequency response functions and operational deflection shapes, two input excitations under prescribed conditions result in tailored traveling waves. The emphasis of this article is the application of projection-based model reduction techniques to scale-down the large-scale FE plate model. Four model reduction techniques are applied and their performances are studied. This article also discusses the stability issues associated with the rigid-body modes. Furthermore, the reduced-order models are utilized to simulate the steady-state frequency and time response of the plate. The results are in agreement with the experimental and the full-scale FE model results.

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Evaluation of staged air and overfire air in regulating air-staging conditions within a large-scale down-fired furnace

Applied Thermal Engineering ◽

10.1016/j.applthermaleng.2014.03.009 ◽

2014 ◽

Vol 67 (1-2) ◽

pp. 97-105 ◽

Cited By ~ 19

Author(s):

Min Kuang ◽

Zhengqi Li ◽

Zhongqian Ling ◽

Xianyang Zeng

Keyword(s):

Large Scale ◽

Scale Down ◽

Air Staging

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A safe, effective, and facility compatible cleaning in place procedure for affinity resin in large-scale monoclonal antibody purification

Journal of Chromatography A ◽

10.1016/j.chroma.2013.07.096 ◽

2013 ◽

Vol 1308 ◽

pp. 86-95 ◽

Cited By ~ 14

Author(s):

Lu Wang ◽

Jill Dembecki ◽

Neil E. Jaffe ◽

Brian W. O’Mara ◽

Hui Cai ◽

...

Keyword(s):

Monoclonal Antibody ◽

Large Scale ◽

Antibody Purification ◽

Affinity Resin

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Development and fine-tuning of a scale down model for process characterization studies of a monoclonal antibody upstream production process

BMC Proceedings ◽

10.1186/1753-6561-5-s8-p70 ◽

2011 ◽

Vol 5 (S8) ◽

Cited By ~ 5

Author(s):

Mareike Harmsen ◽

Jimmy Stofferis ◽

Laetitia Malphettes

Keyword(s):

Monoclonal Antibody ◽

Production Process ◽

Fine Tuning ◽

Process Characterization ◽

Scale Down ◽

Characterization Studies

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Relationship between IL10 and PD-L1 in Liver Hepatocellular Carcinoma Tissue and Cell Lines

BioMed Research International ◽

10.1155/2020/8910183 ◽

2020 ◽

Vol 2020 ◽

pp. 1-14

Author(s):

Qian Qian ◽

Changping Wu ◽

Jianping Chen ◽

Weibing Wang

Keyword(s):

Hepatocellular Carcinoma ◽

Monoclonal Antibody ◽

Cell Lines ◽

Western Blotting ◽

Large Scale ◽

Expression Level ◽

Survival Duration ◽

Great Promise ◽

Clinical Response Rate ◽

Liver Hepatocellular Carcinoma

Background. Despite the large-scale clinical application of programmed death-ligand 1 (PD-L1) monoclonal antibody, reduction in its clinical response rate has become a gradual problem. As such, use of PD-L1 monoclonal antibody in combination with other anticarcinoma drugs has been the main strategy in improving its efficacy. Interleukin 10 (IL10) is a recognized inflammatory and immunosuppressive factor. Previous studies have suggested that there is a link between PD-L1 and IL10. Objective. This study was aimed at clarifying the relationship between PD-L1 and IL10 in liver hepatocellular carcinoma (LIHC) and whether IL10 enhances the efficacy of PD-L1 inhibitor. Methods. Expression levels of PD-L1 and IL10 in carcinoma and adjacent tissues were tested by immunochemistry, Western blotting, and RT-PCR. Survival duration and follow-up data of each patient were recorded. LIHC cell lines Bel7405 and MHCC 97-H were used for in vitro experiments. Exogenous IL10 and anti-IL10 were added to cell supernatant. Expression level of PD-L1 in the LIHC cell lines was determined using Western blotting and ELISA. CCK8 and transwell assays were adopted to examine the effect of PD-L1 combined with IL10 on proliferation, invasion, and metastasis of LIHC cells. Results. The survival period of patients with low expression of IL10 was longer than that of patients with high expression (P=0.01). Overexpression of PD-L1 increased the IL10 and Met levels in LIHC tissues and cell lines. IL10 downregulated the expression level of PD-L1 and enhanced the efficacy of crizotinib via the Met signaling pathway in the LIHC cells. Conclusions. A combination of IL10 and PD-L1 inhibitor holds great promise as an effective treatment for LIHC.

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