Abstract:
:Therapeutic monoclonal antibodies (mAbs) are highly complex proteins that must be exhaustively characterized according to the regulatory authorities' recommendations. MAbs display micro-heterogeneity mainly due to their post-translational modifications, but also to their susceptibility to chemical and physical degradations. Among these degradations, aggregation is quite frequent, initiated by protein denaturation and then dimer formation. Here, we investigated the nature and structure of the high molecular weight species (HMW) present at less than 1% in an unstressed formulated roledumab biopharmaceutical, as a model of high purity mAb. HMW species were first purified through preparative size-exclusion chromatography (SEC) and then analyzed by a combination of chromatographic methods (ion-exchange chromatography (IEX), SEC) coupled to native mass spectrometry (MS), as well as sodium dodecyl sulfate-polyacrylamide gel electrophoresis and capillary gel electrophoresis under non-reducing conditions. Both covalently and non-covalently bound dimers were identified at a proportion of 50/50. In-depth characterization of the HMW fraction by SEC and IEX hyphenated to native MS revealed the presence of three mAb dimer forms having the same mass, but differing by their charge and size. They were attributed to different compact and elongated dimers. Finally, high-resolution middle-up approaches using different enzymes (IdeS and IgdE) were performed to determine the mAb domains implicated in the dimerization. Our results revealed that the roledumab dimers were associated mainly by a single Fab-to-Fab arm-bound association.
journal_name
MAbsjournal_title
mAbsauthors
Rouby G,Tran NT,Leblanc Y,Taverna M,Bihoreau Ndoi
10.1080/19420862.2020.1781743subject
Has Abstractpub_date
2020-01-01 00:00:00pages
e1781743issue
1eissn
1942-0862issn
1942-0870journal_volume
12pub_type
杂志文章相关文献
mAbs文献大全abstract::Therapeutic antibodies continue to develop as an emerging drug class, with a need for preclinical tools to better predict in vivo characteristics. Transgenic mice expressing human neonatal Fc receptor (hFcRn) have potential as a preclinical pharmacokinetic (PK) model to project human PK of monoclonal antibodies (mAbs)...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2016.1193660
更新日期:2016-08-01 00:00:00
abstract::Multispecific antibody formats provide a promising platform for the development of novel therapeutic concepts that could facilitate the generation of safer, more effective pharmaceuticals. However, the production and use of such antibody-based multispecifics is often made complicated by: 1) the instability of the anti...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2016.1248012
更新日期:2017-01-01 00:00:00
abstract::While many antibody therapeutics are formulated at low concentration (~10-20 mg/mL) for intravenous administration, high concentration (> 100 mg/mL) formulations may be required for subcutaneous delivery in certain clinical indications. For such high concentration formulations, product color is more apparent due to th...
journal_title:mAbs
pub_type: 杂志文章
doi:10.4161/mabs.28257
更新日期:2014-05-01 00:00:00
abstract::Neutralizing antibody responses to the surface glycoproteins of enveloped viruses play an important role in immunity. Many of these glycoproteins, including the severe acute respiratory syndrome-coronavirus (SARS-CoV) spike (S) protein form trimeric units in the membrane of the native virion. There is substantial expe...
journal_title:mAbs
pub_type: 杂志文章
doi:10.4161/mabs.2.1.10788
更新日期:2010-01-01 00:00:00
abstract::Bispecific antibodies can uniquely influence cellular responses, but selecting target combinations for optimal functional activity remains challenging. Here we describe a high-throughput, combinatorial, phenotypic screening approach using a new bispecific antibody target discovery format, allowing screening of hundred...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2020.1859049
更新日期:2021-01-01 00:00:00
abstract::Antibody-drug conjugates (ADCs) represent a new class of cancer therapeutics. Their design involves a tumor-specific antibody, a linker and a cytotoxic payload. They were designed to allow specific targeting of highly potent cytotoxic agents to tumor cells whilst sparing normal cells. Frequent toxicities that may be d...
journal_title:mAbs
pub_type: 杂志文章,评审
doi:10.1080/19420862.2016.1156829
更新日期:2016-05-01 00:00:00
abstract::A costimulatory signal is required for the full activation of T cells, in addition to the antigen-specific signal via the T cell receptor. The inducible costimulator, ICOS is one of the costimulatory molecules that play an essential role in this process, particularly in the expansion or the development of effector T c...
journal_title:mAbs
pub_type: 杂志文章
doi:10.4161/mabs.1.5.9633
更新日期:2009-09-01 00:00:00
abstract::Ricin is a toxin that could potentially be used as a bioweapon. We identified anti-ricin A chain antibodies by sequencing the antibody repertoire from immunized mice and by selecting high affinity antibodies using yeast surface display. These methods led to the isolation of multiple antibodies with high (sub-nanomolar...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2016.1190059
更新日期:2016-08-01 00:00:00
abstract::Ectodomains of target antigens for antibody-based therapies can be shed from the target cell surface and found in sera of patients. Shed ectodomains of therapeutic targets not only pose the risk of sequestering therapeutic antibodies but, in a multimeric form, of triggering T cell activation by bispecific antibodies b...
journal_title:mAbs
pub_type: 杂志文章
doi:10.4161/mabs.3.1.14193
更新日期:2011-01-01 00:00:00
abstract::The advantages of site-specific over stochastic bioconjugation technologies include homogeneity of product, minimal perturbation of protein structure/function, and - increasingly - the ability to perform structure activity relationship studies at the conjugate level. When selecting the optimal location for site-specif...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2018.1512327
更新日期:2018-11-01 00:00:00
abstract::Highly concentrated antibody solutions often exhibit high viscosities, which present a number of challenges for antibody-drug development, manufacturing and administration. The antibody sequence is a key determinant for high viscosity of highly concentrated solutions; therefore, a sequence- or structure-based tool tha...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2015.1099773
更新日期:2016-01-01 00:00:00
abstract::Tandem single-chain variable fragment (scFv) bispecific antibodies (bsAb) are one of the most promising bsAb formats reported thus far. Yet, because of their increased aggregation propensity, high impurity content due to low expression level, smaller size and lack of the Fc region, it is challenging to isolate these p...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2020.1718440
更新日期:2020-01-01 00:00:00
abstract::Process intensification has shown great potential to increase productivity and reduce costs in biomanufacturing. This case study describes the evolution of a manufacturing process from a conventional processing scheme at 1000-L scale (Process A, n = 5) to intensified processing schemes at both 1000-L (Process B, n = 8...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2020.1770669
更新日期:2020-01-01 00:00:00
abstract::Efficient production of large quantities of therapeutic antibodies is becoming a major goal of the pharmaceutical industry. We developed a proprietary expression system using a polyprotein precursor-based approach to antibody expression in mammalian cells. In this approach, the coding regions for heavy and light chain...
journal_title:mAbs
pub_type: 杂志文章
doi:10.4161/mabs.25161
更新日期:2013-07-01 00:00:00
abstract::Antibodies provide immune protection by recognizing antigens of diverse chemical properties, but elucidating the amino acid sequence-function relationships underlying the specificity and affinity of antibody-antigen interactions remains challenging. We designed and constructed phage-displayed synthetic antibody librar...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2018.1550320
更新日期:2019-02-01 00:00:00
abstract::Therapeutic monoclonal antibodies (mAbs) currently dominate the biologics marketplace. Development of a new therapeutic mAb candidate is a complex, multistep process and early stages of development typically begin in an academic research environment. Recently, a number of facilities and initiatives have been launched ...
journal_title:mAbs
pub_type: 杂志文章,评审
doi:10.4161/mabs.3.5.16968
更新日期:2011-09-01 00:00:00
abstract::Biosimilars are designed to be highly similar to approved or licensed (reference) biologics and are evaluated based on the totality of evidence from extensive analytical, nonclinical and clinical studies. As part of the stepwise approach recommended by regulatory agencies, the first step in the clinical evaluation of ...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2016.1171431
更新日期:2016-07-01 00:00:00
abstract::Due to their excellent specificity for a single epitope, monoclonal antibodies (mAbs) present a means of influencing the function of cells at the molecular level. In particular they show great promise in the treatment of cancer because they can inhibit cancer cell proliferation, tumor angiogenesis, invasiveness and ma...
journal_title:mAbs
pub_type: 杂志文章,评审
doi:10.4161/mabs.3.5.16089
更新日期:2011-09-01 00:00:00
abstract::Animal cell culture technology has advanced significantly over the last few decades and is now generally considered a reliable, robust and relatively mature technology. A range of biotherapeutics are currently synthesized using cell culture methods in large scale manufacturing facilities that produce products for both...
journal_title:mAbs
pub_type: 杂志文章,评审
doi:10.4161/mabs.2.5.12720
更新日期:2010-09-01 00:00:00
abstract::Recombinant antibody single-chain variable fragments (scFv) are difficult to purify homogeneously from a protein complex mixture. The most effective, specific and fastest method of purification is an affinity chromatography on Protein L (PpL) matrix. This protein is a multi-domain bacterial surface protein that is abl...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2015.1116657
更新日期:2016-01-01 00:00:00
abstract::Genetically encoded fluorescent antibodies are desirable for many applications in biotechnology, proteomics, microscopy, cell biology and molecular diagnostics, although efficient production of fluorescent IgGs in mammalian cells has been hampered by different and mutually incompatible secretion- and folding-requireme...
journal_title:mAbs
pub_type: 杂志文章
doi:10.4161/mabs.2.6.13179
更新日期:2010-11-01 00:00:00
abstract::The neonatal Fc receptor (FcRn) is important for the metabolic fate of IgG antibodies in vivo. Analysis of the interaction between FcRn and IgG in vitro might provide insight into the structural and functional integrity of therapeutic IgG that may affect pharmacokinetics (PK) in vivo. We developed a standardized pH gr...
journal_title:mAbs
pub_type: 杂志文章
doi:10.4161/mabs.24981
更新日期:2013-07-01 00:00:00
abstract::Monoclonal antibody (mAb) candidates from high-throughput screening or binding affinity optimization often contain mutations leading to liabilities for further development of the antibody, such as aggregation-prone regions and lack of solubility. In this work, we optimized a candidate integrin α11-binding mAb for deve...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2018.1475871
更新日期:2018-08-01 00:00:00
abstract::Monoclonal antibodies are used with great success in many different therapeutic domains. In order to satisfy the growing demand and to lower the production cost of these molecules, many alternative systems have been explored. Among them, the baculovirus/insect cells system is a good candidate. This system is very safe...
journal_title:mAbs
pub_type: 杂志文章,评审
doi:10.4161/mabs.19942
更新日期:2012-05-01 00:00:00
abstract::The development of alternative therapeutic strategies to tumor necrosis factor (TNF)-blocking antibodies for the treatment of inflammatory diseases has generated increasing interest. In particular, selective inhibition of TNF receptor 1 (TNFR1) promises a more precise intervention, tackling only the pro-inflammatory r...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2019.1596512
更新日期:2019-05-01 00:00:00
abstract::The development and production of recombinant monoclonal antibodies is well established. Although most of these are IgGs, there is also great interest in producing recombinant IgAs since this isotype plays a critical role in providing immunologic protection at mucosal surfaces. The choice of expression system for prod...
journal_title:mAbs
pub_type: 杂志文章
doi:10.4161/mabs.2.3.11802
更新日期:2010-05-01 00:00:00
abstract::The Fc glycosylation of therapeutic antibodies is crucial for their effector functions and their behavior in pharmacokinetics and pharmacodynamics. To monitor the Fc glycosylation in bioprocess development and characterization,high-throughput techniques for glycosylation analysis are needed. Here, we describe the deve...
journal_title:mAbs
pub_type: 杂志文章
doi:10.4161/mabs.26712
更新日期:2014-01-01 00:00:00
abstract::Severe forms of dengue virus (DENV) infection frequently cause high case fatality rate. Currently, there is no effective vaccine against the infection. Clinical cases are given only palliative treatment as specific anti-DENV immunotherapy is not available and it is urgently required. In this study, human single-chain ...
journal_title:mAbs
pub_type: 杂志文章
doi:10.4161/mabs.27874
更新日期:2014-03-01 00:00:00
abstract::Glycosylation is a critical attribute for development and manufacturing of therapeutic monoclonal antibodies (mAbs) in the pharmaceutical industry. Conventional antibody glycan analysis is usually achieved by the 2-aminobenzamide (2-AB) hydrophilic interaction liquid chromatography (HILIC) method following the release...
journal_title:mAbs
pub_type: 杂志文章
doi:10.1080/19420862.2016.1156828
更新日期:2016-05-01 00:00:00
abstract::The neonatal Fc receptor (FcRn) plays an important and well-known role in antibody recycling in endothelial and hematopoietic cells and thus it influences the systemic pharmacokinetics (PK) of immunoglobulin G (IgG). However, considerably less is known about FcRn's role in the metabolism of IgG within individual tissu...
journal_title:mAbs
pub_type: 杂志文章
doi:10.4161/mabs.28254
更新日期:2014-05-01 00:00:00