In vitro functional characterization of biosimilar therapeutic antibodies

Láng, Júlia Anna; Balogh, Zsófia Cselovszkiné; Nyitrai, Mónika Fizilné; Juhasz, C.C.; Gilicze, Anna Katalin Baráné; Iliás, Attila; Zólyomi, Zsolt; Bödör, Csaba; Rábai, Erzsébet

doi:10.1016/j.ddtec.2020.11.010

Cited by 8 publications

(5 citation statements)

References 53 publications

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“…The binding affinities of the scFv and LP-scFv to HER2 were measured using label-free biolayer interferometry (BLI) [ 28 , 29 ], as shown in Figure S2 . Table 1 presents the calculated association ( k on ) and dissociation ( k off ) rate constants, along with the equilibrium dissociation constants ( K D = k off / k on ).…”

Section: Resultsmentioning

confidence: 99%

A Cell-Penetrating Peptide Improves Anti-HER2 Single-Chain Variable Fragment Internalization and Antitumor Activity against HER2-Positive Breast Cancer In Vitro and In Vivo

Li,

Zhou,

et al. 2024

Molecules

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Cell-penetrating peptides (CPPs) are invaluable tools for delivering various substances into cells by crossing biological membranes. However, the effects of cell-penetrating peptide fusion proteins on the biological activity of antibodies remain to be fully understood. Here, we engineered a recombinant protein, LP-scFv, which combines the single-chain variable region of anti-human epidermal growth factor receptor-2 with a novel and non-oxic cell-penetrating peptide as a leader peptide. The introduction of this leader peptide led to a more than twofold increase in the internalization efficiency of the single-chain antibody, as confirmed using microscopic analysis and flow cytometry. The effects of the single-chain antibodies and LP-scFv on cell viability were evaluated using the MTT assay. Both the single-chain antibodies and LP-scFv reduced the viability of BT474 and NCI-N87 cells in a dose-dependent manner while exhibiting minimal toxicity towards MCF-7 and MCF-10A cells. Further investigation into LP-scFv’s mechanism revealed that the induced leader peptide does not alter the MAPK-ERK1/2 and PI3K/AKT pathways of single-chain antibodies. An enhanced antitumor activity was also confirmed in an NCI-N87 tumor xenograft model in mice with a reduction of 45.2% in tumor growth inhibition (vs. 23.1% for scFv) with a 50 mg/kg dose after orthotopic injection administration, which was equivalent to that of trastuzumab (vs. 55.7% for trastuzumab). Overall, these results indicate that LP-scFv exhibits significant permeation activity in HER2-positive cells to enhance the intracellular dose effect on antitumor activity in vitro and in vivo. This research lays the foundation for designing novel antibody-based therapies for cancer.

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Section: Resultsmentioning

confidence: 99%

A Cell-Penetrating Peptide Improves Anti-HER2 Single-Chain Variable Fragment Internalization and Antitumor Activity against HER2-Positive Breast Cancer In Vitro and In Vivo

Li,

Zhou,

et al. 2024

Molecules

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“…On the other hand, functional tests related to the possible MOA, such as apoptosis, complement-dependent cytotoxicity, antibody-dependent cellular phagocytosis, and antibody-dependent cellular cytotoxicity, among others, are necessary but not essential, especially when it is not relevant. For instance, functional tests (ADCC, ADCP, and CDC) are unnecessary for a product that predominantly targets a soluble antigen [165][166][167][168][169].…”

Section: Function Testingmentioning

confidence: 99%

Reinventing Therapeutic Proteins: Mining a Treasure of New Therapies

Niazi

Mariam

2023

Biologics

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Reinventing approved therapeutic proteins for a new dose, a new formulation, a new route of administration, an improved safety profile, a new indication, or a new conjugate with a drug or a radioactive source is a creative approach to benefit from the billions spent on developing new therapeutic proteins. These new opportunities were created only recently with the arrival of AI/ML tools and high throughput screening technologies. Furthermore, the complex nature of proteins offers mining opportunities that are not possible with chemical drugs; bringing in newer therapies without spending billions makes this path highly lucrative financially while serving the dire needs of humanity. This paper analyzes several practical reinventing approaches and suggests regulatory strategies to reduce development costs significantly. This should enable the entry of hundreds of new therapies at affordable costs.

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“…Для оценки биологической активности препаратов МкАТ применяют в основном методики in vitro на культурах клеток, отражающие механизм их действия в организме человека, что позволяет оценить взаимодействие МкАТ с соответствующими мишенями [6,9,18]. На рисунке 1 представлены основные механизмы действия терапевтических МкАТ 4 .…”

Section: особенности оценки биологических свойств препаратов моноклон...unclassified

“…МСО Всемирной организации здравоохранения (ВОЗ) являются первичными СО и доступны для широкого круга действующих веществ лекарственных препаратов, включая БтЛП 8 . Кандидаты в МСО производятся в соответствии с определенными стандартными процедурами, которые оптимизируют сохранение биологической активности и других важных характеристик образцов для повышения их стабильности 9 . После подтверждения пригодности кандидата в рамках международного совместного исследования он утверждается 7 https://www.ema.europa.eu/en/documents/presentation/session-1-cmc-innovator-industry-presentation-prof-georg-b-kresse_en.pdf 8 Providing international biological reference preparations.…”

Section: необходимость разработки международных стандартных образцов ...unclassified

“…с референтным препаратом (РП) 2 [2]. Оценка фармакодинамических свойств и биологической активности МкАТ важна для понимания структурно-функциональных особенностей биоаналогичных препаратов, механизмов их взаимодействия с биологическими мишенями, что формирует основу для установления функциональной сопоставимости с РП [8,9].…”

Section: Introductionunclassified

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International standards for monoclonal antibodies for assessing the biological activity of medicines: A status update

Gayderova,

Alpatova,

Lysikova

et al. 2023

Biological Products. Prevention, Diagnosis, Treatment

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Scientific relevance. The clinical effects and the expiration of patents for original (reference) biotechnological medicines based on monoclonal antibodies (mAbs) stimulated the development of biosimilar mAbs. The quality profile of a biosimilar mAb should correspond to the quality of the reference medicinal product. When demonstrating biosimilarity and determining the activity of medicines as part of batch quality control, analysts should study the biological properties of mAbs using suitable reference standards. The lack of international standards (ISs) makes mAb manufacturers use in-house reference standards. There is a risk of obtaining non-uniform quality and efficacy data because of the use of in-house reference standards, the heterogeneity and structural complexity of mAbs, and the relationship between the biological activity and efficacy of mAbs.Aim. This study aimed to analyse the relevance of and need for ISs for the biological activity of biotherapeutic mAbs and to define the role of reference medicinal products and ISs in assessing biosimilarity and testing medicines throughout their lifecycle.Discussion. This review covers the issues arising from the lack of ISs for assessing the biological activity of mAbs and the role and significance of reference products and ISs for biosimilars. The authors describe the specifics of studying the biological properties of mAbs and summarise the data on the need to develop and use ISs for the standardisation of biological tests. This review presents the results of studies on the first ISs established by the World Health Organisation to assess the biological activity of mAbs; these results suggest the need to standardise mAbs using ISs to ensure the quality, safety, and efficacy of mAb therapy.Conclusions. The use of ISs for mAbs plays a key role in harmonising biological activity assessments. Publicly available ISs serve as primary standards for the calibration of secondary reference materials. Moreover, ISs are required for the harmonisation of activity evaluation (in IU) between laboratories and for the consistency of the activity of various medicinal products from different manufacturers that share the same INN. The use of ISs by mAb manufacturers will contribute to ensuring the quality of mAbs and clinical monitoring of the effectiveness of their use.

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In vitro functional characterization of biosimilar therapeutic antibodies

Cited by 8 publications

References 53 publications

A Cell-Penetrating Peptide Improves Anti-HER2 Single-Chain Variable Fragment Internalization and Antitumor Activity against HER2-Positive Breast Cancer In Vitro and In Vivo

A Cell-Penetrating Peptide Improves Anti-HER2 Single-Chain Variable Fragment Internalization and Antitumor Activity against HER2-Positive Breast Cancer In Vitro and In Vivo

Reinventing Therapeutic Proteins: Mining a Treasure of New Therapies

International standards for monoclonal antibodies for assessing the biological activity of medicines: A status update

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