PDADMAC flocculation of Chinese hamster ovary cells: enabling a centrifuge-less harvest process for monoclonal antibodies.

Abstract:

:High titer (>10 g/L) monoclonal antibody (mAb) cell culture processes are typically achieved by maintaining high viable cell densities over longer culture durations. A corresponding increase in the solids and sub-micron cellular debris particle levels are also observed. This higher burden of solids (≥15%) and sub-micron particles typically exceeds the capabilities of a continuous centrifuge to effectively remove the solids without a substantial loss of product and/or the capacity of the harvest filtration train (depth filter followed by membrane filter) used to clarify the centrate. We discuss here the use of a novel and simple two-polymer flocculation method used to harvest mAb from high cell mass cell culture processes. The addition of the polycationic polymer, poly diallyldimethylammonium chloride (PDADMAC) to the cell culture broth flocculates negatively-charged cells and cellular debris via an ionic interaction mechanism. Incorporation of a non-ionic polymer such as polyethylene glycol (PEG) into the PDADMAC flocculation results in larger flocculated particles with faster settling rate compared to PDADMAC-only flocculation. PDADMAC also flocculates the negatively-charged sub-micron particles to produce a feed stream with a significantly higher harvest filter train throughput compared to a typical centrifuged harvest feed stream. Cell culture process variability such as lactate production, cellular debris and cellular densities were investigated to determine the effect on flocculation. Since PDADMAC is cytotoxic, purification process clearance and toxicity assessment were performed.

journal_name

MAbs

journal_title

mAbs

authors

McNerney T,Thomas A,Senczuk A,Petty K,Zhao X,Piper R,Carvalho J,Hammond M,Sawant S,Bussiere J

doi

10.1080/19420862.2015.1007824

subject

Has Abstract

pub_date

2015-01-01 00:00:00

pages

413-28

issue

2

eissn

1942-0862

issn

1942-0870

journal_volume

7

pub_type

杂志文章

相关文献

mAbs文献大全
  • Lepidopteran cells, an alternative for the production of recombinant antibodies?

    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

    authors: Cérutti M,Golay J

    更新日期:2012-05-01 00:00:00

  • Investigation of the effect of salt additives in Protein L affinity chromatography for the purification of tandem single-chain variable fragment bispecific antibodies.

    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

    authors: Chen SW,Tan D,Yang YS,Zhang W

    更新日期:2020-01-01 00:00:00

  • Generation of human single-chain variable fragment antibodies specific to dengue virus non-structural protein 1 that interfere with the virus infectious cycle.

    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

    authors: Poungpair O,Bangphoomi K,Chaowalit P,Sawasdee N,Saokaew N,Choowongkomon K,Chaicumpa W,Yenchitsomanus PT

    更新日期:2014-03-01 00:00:00

  • Quantitative cumulative biodistribution of antibodies in mice: effect of modulating binding affinity to the neonatal Fc receptor.

    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

    authors: Yip V,Palma E,Tesar DB,Mundo EE,Bumbaca D,Torres EK,Reyes NA,Shen BQ,Fielder PJ,Prabhu S,Khawli LA,Boswell CA

    更新日期:2014-05-01 00:00:00

  • Continuous bioprocessing: the real thing this time? 10(th) Annual bioProcessUK Conference, December 3-4, 2013, London, UK.

    abstract::The Annual bioProcessUK Conference has acted as the key networking event for bioprocess scientists and engineers in the UK for the past 10 years. The following article is a report from the sessions that focused on continuous bioprocessing during the 10(th) Annual bioProcessUK Conference (London, December 2013). These ...

    journal_title:mAbs

    pub_type:

    doi:10.4161/mabs.36151

    authors: Farid SS,Thompson B,Davidson A

    更新日期:2014-01-01 00:00:00

  • A chemical and computational approach to comprehensive glycation characterization on antibodies.

    abstract::Non-enzymatic glycation is a challenging post-translational modification to characterize due to the structural heterogeneity it generates in proteins. Glycation has become increasingly recognized as an important product quality attribute to monitor, particularly for the biotechnology sector, which produces recombinant...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.1080/19420862.2015.1046663

    authors: Saleem RA,Affholter BR,Deng S,Campbell PC,Matthies K,Eakin CM,Wallace A

    更新日期:2015-01-01 00:00:00

  • Approaches to improve tumor accumulation and interactions between monoclonal antibodies and immune cells.

    abstract::Monoclonal antibodies (mAb) have become a mainstay in tumor therapy. Clinical responses to mAb therapy, however, are far from optimal, with many patients presenting native or acquired resistance or suboptimal responses to a mAb therapy. MAbs exert antitumor activity through different mechanisms of action and we propos...

    journal_title:mAbs

    pub_type: 杂志文章,评审

    doi:10.4161/mabs.22775

    authors: Marcucci F,Bellone M,Rumio C,Corti A

    更新日期:2013-01-01 00:00:00

  • Target-independent variable region mediated effects on antibody clearance can be FcRn independent.

    abstract::The importance of the neonatal Fc receptor (FcRn) in extending the serum half-life of monoclonal antibodies (mAbs) is well demonstrated, and has led to the development of multiple engineering approaches designed to alter Fc interactions with FcRn. Recent reports have additionally highlighted the effect of nonspecific ...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.1080/19420862.2016.1208330

    authors: Kelly RL,Yu Y,Sun T,Caffry I,Lynaugh H,Brown M,Jain T,Xu Y,Wittrup KD

    更新日期:2016-10-01 00:00:00

  • Dual targeting strategies with bispecific antibodies.

    abstract::Monoclonal antibodies are widely used for the treatment of cancer, inflammatory and infectious diseases and other disorders. Most of the marketed antibodies are monospecific and therefore capable of interacting and interfering with a single target. However, complex diseases are often multifactorial in nature, and invo...

    journal_title:mAbs

    pub_type: 杂志文章,评审

    doi:10.4161/mabs.4.2.19000

    authors: Kontermann RE

    更新日期:2012-03-01 00:00:00

  • Rapid identification of highly potent human anti-GPCR antagonist monoclonal antibodies.

    abstract::Complex cellular targets such as G protein-coupled receptors (GPCRs), ion channels, and other multi-transmembrane proteins represent a significant challenge for therapeutic antibody discovery, primarily because of poor stability of the target protein upon extraction from cell membranes. To assess whether a limited set...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.1080/19420862.2020.1755069

    authors: Scott MJ,Jowett A,Orecchia M,Ertl P,Ouro-Gnao L,Ticehurst J,Gower D,Yates J,Poulton K,Harris C,Mullin MJ,Smith KJ,Lewis AP,Barton N,Washburn ML,de Wildt R

    更新日期:2020-01-01 00:00:00

  • Fluorescent Citrine-IgG fusion proteins produced in mammalian cells.

    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

    authors: Haas AK,von Schwerin C,Matscheko D,Brinkmann U

    更新日期:2010-11-01 00:00:00

  • Thermodynamic and structural characterization of an antibody gel.

    abstract::Although extensively studied, protein-protein interactions remain highly elusive and are of increasing interest in drug development. We show the assembly of a monoclonal antibody, using multivalent carboxylate ions, into highly-ordered structures. While the presence and function of similar structures in vivo are not k...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.4161/mabs.23183

    authors: Esue O,Xie AX,Kamerzell TJ,Patapoff TW

    更新日期:2013-03-01 00:00:00

  • Sequencing antibody repertoires: the next generation.

    abstract::Genomic studies have been revolutionized by the use of next generation sequencing (NGS) that delivers huge amounts of sequence information in a short span of time. The number of applications for NGS is rapidly expanding and significantly transforming many areas of life sciences. The field of antibody research and disc...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.4161/mabs.3.1.14169

    authors: Fischer N

    更新日期:2011-01-01 00:00:00

  • Concentrations of EpCAM ectodomain as found in sera of cancer patients do not significantly impact redirected lysis and T-cell activation by EpCAM/CD3-bispecific BiTE antibody MT110.

    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

    authors: Petsch S,Gires O,Rüttinger D,Denzel S,Lippold S,Baeuerle PA,Wolf A

    更新日期:2011-01-01 00:00:00

  • Antibodies to watch in 2010.

    abstract::Monoclonal antibodies (mAbs) are a burgeoning class of therapeutics, with more than 25 approved in countries worldwide. Novel molecules are entering clinical study at a rate of nearly 40 per year, and the commercial pipeline includes approximately 240 mAb therapeutics in clinical studies that have not yet progressed t...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.4161/mabs.2.1.10677

    authors: Reichert JM

    更新日期:2010-01-01 00:00:00

  • Evaluating imbalances of adverse events during biosimilar development.

    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

    authors: Vana AM,Freyman AW,Reich SD,Yin D,Li R,Anderson S,Jacobs IA,Zacharchuk CM,Ewesuedo R

    更新日期:2016-07-01 00:00:00

  • Phage display-based generation of novel internalizing antibody fragments for immunotoxin-based treatment of acute myeloid leukemia.

    abstract::The current standard treatment for acute myeloid leukemia (AML) is chemotherapy based on cytarabine and daunorubicine (7 + 3), but it discriminates poorly between malignant and benign cells. Dose-limiting off‑target effects and intrinsic drug resistance result in the inefficient eradication of leukemic blast cells and...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.1080/19420862.2015.1007818

    authors: Fitting J,Blume T,Ten Haaf A,Blau W,Gattenlöhner S,Tur MK,Barth S

    更新日期:2015-01-01 00:00:00

  • Antibody-drug conjugates targeting TROP-2 and incorporating SN-38: A case study of anti-TROP-2 sacituzumab govitecan.

    abstract::Antibody-drug conjugates (ADCs) that exploit the active metabolite SN-38, which is derived from the popular anticancer drug, irinotecan (a camptothecin that inhibits the nuclear topoisomerase I enzyme, inducing double-stranded DNA breaks during the mitotic S-phase of affected cells), represent a substantial advance in...

    journal_title:mAbs

    pub_type: 杂志文章,评审

    doi:10.1080/19420862.2019.1632115

    authors: Goldenberg DM,Sharkey RM

    更新日期:2019-08-01 00:00:00

  • Cysteinylation of a monoclonal antibody leads to its inactivation.

    abstract::Post-translational modifications can have a signification effect on antibody stability. A comprehensive approach is often required to best understand the underlying reasons the modification affects the antibody's potency or aggregation state. Monoclonal antibody 001 displayed significant variation in terms of potency,...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.1080/19420862.2016.1160179

    authors: McSherry T,McSherry J,Ozaeta P,Longenecker K,Ramsay C,Fishpaugh J,Allen S

    更新日期:2016-05-01 00:00:00

  • Analytical FcRn affinity chromatography for functional characterization of monoclonal antibodies.

    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

    authors: Schlothauer T,Rueger P,Stracke JO,Hertenberger H,Fingas F,Kling L,Emrich T,Drabner G,Seeber S,Auer J,Koch S,Papadimitriou A

    更新日期:2013-07-01 00:00:00

  • Rapid identification of an antibody DNA construct rearrangement sequence variant by mass spectrometry.

    abstract::During cell line development for an IgG1 antibody candidate (mAb1), a C-terminal extension was identified in 2 product candidate clones expressed in CHO-K1 cell line. The extension was initially observed as the presence of anomalous new peaks in these clones after analysis by cation exchange chromatography (CEX-HPLC) ...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.4161/mabs.36222

    authors: Scott RA,Rogers R,Balland A,Brady LJ

    更新日期:2014-01-01 00:00:00

  • Metal ion interactions with mAbs: Part 1.

    abstract::Fragmentation in the hinge region of an IgG1 monoclonal antibody (mAb) can affect product stability, potentially causing changes in potency and efficacy. Metals ions, such as Cu(2+), can bind to the mAb and undergo hydrolysis or oxidation, which can lead to cleavage of the molecule. To better understand the mechanism ...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.1080/19420862.2015.1062193

    authors: Glover ZK,Basa L,Moore B,Laurence JS,Sreedhara A

    更新日期:2015-01-01 00:00:00

  • Going native: Direct high throughput screening of secreted full-length IgG antibodies against cell membrane proteins.

    abstract::Gel microdroplet - fluorescence activated cell sorting (GMD-FACS) is an innovative high throughput screening platform for recombinant protein libraries, and we show here that GMD-FACS can overcome many of the limitations associated with conventional screening methods for antibody libraries. For example, phage and cell...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.1080/19420862.2017.1381812

    authors: Fang Y,Chu TH,Ackerman ME,Griswold KE

    更新日期:2017-11-01 00:00:00

  • Molecular characterization of the cervical and systemic B-cell repertoire: Unique, yet overlapping, immune compartments of an HIV-1 resistant individual.

    abstract::The cervical mucosa of women who are highly exposed to HIV-1, yet remain persistently seronegative (HEPS), presents a unique opportunity to study the dynamics of an immune compartment potentially capable of preventing HIV-1 infection. Herein, we provide a detailed characterization of the immunoglobulin repertoire of c...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.4161/mabs.3.2.14858

    authors: Gaudet RG,Breden F,Plummer F,Berry JD

    更新日期:2011-03-01 00:00:00

  • Antibody-drug conjugates: Design and development for therapy and imaging in and beyond cancer, LabEx MAbImprove industrial workshop, July 27-28, 2017, Tours, France.

    abstract::The annual "Antibody Industrial Symposium", co organized by LabEx MAbImprove, MabDesign and Polepharma, was held in Tours, France on June 27-28, 2017. The focus was on antibody-drug-conjugates (ADCs), new entities which realize the hope of Paul Ehrlich's magic bullet. ADCs result from the bioconjugation of a highly cy...

    journal_title:mAbs

    pub_type:

    doi:10.1080/19420862.2017.1412130

    authors: Martin C,Kizlik-Masson C,Pèlegrin A,Watier H,Viaud-Massuard MC,Joubert N

    更新日期:2018-02-01 00:00:00

  • IgE immunotherapy: a novel concept with promise for the treatment of cancer.

    abstract::The importance of antibodies in activating immune responses against tumors is now better appreciated with the emergence of checkpoint blockade antibodies and with engineered antibody Fc domains featuring enhanced capacity to focus potent effector cells against cancer cells. Antibodies designed with Fc regions of the I...

    journal_title:mAbs

    pub_type: 杂志文章,评审

    doi:10.4161/mabs.27029

    authors: Josephs DH,Spicer JF,Karagiannis P,Gould HJ,Karagiannis SN

    更新日期:2014-01-01 00:00:00

  • High throughput detection of antibody self-interaction by bio-layer interferometry.

    abstract::Self-interaction of an antibody may lead to aggregation, low solubility or high viscosity. Rapid identification of highly developable leads remains challenging, even though progress has been made with the introduction of techniques such as self-interaction chromatography (SIC) and cross-interaction chromatography (CIC...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.4161/mabs.26186

    authors: Sun T,Reid F,Liu Y,Cao Y,Estep P,Nauman C,Xu Y

    更新日期:2013-11-01 00:00:00

  • Bispecific antibody target pair discovery by high-throughput phenotypic screening using in vitro combinatorial Fab libraries.

    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

    authors: Bhatta P,Whale KD,Sawtell AK,Thompson CL,Rapecki SE,Cook DA,Twomey BM,Mennecozzi M,Starkie LE,Barry EMC,Peters SJ,Kamal AM,Finney HM

    更新日期:2021-01-01 00:00:00

  • Diabody-Ig: a novel platform for the generation of multivalent and multispecific antibody molecules.

    abstract::Multivalent mono- or bispecific antibodies are of increasing interest for therapeutic applications, such as efficient receptor clustering and activation, or dual targeting approaches. Here, we present a novel platform for the generation of Ig-like molecules, designated diabody-Ig (Db-Ig). The antigen-binding site of D...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.1080/19420862.2019.1603024

    authors: Seifert O,Rau A,Beha N,Richter F,Kontermann RE

    更新日期:2019-07-01 00:00:00

  • A novel IgE-neutralizing antibody for the treatment of severe uncontrolled asthma.

    abstract::The critical role played by IgE in allergic asthma is well-documented and clinically precedented, but some patients in whom IgE neutralization may still offer clinical benefit are excluded from treatment with the existing anti-IgE therapy, omalizumab, due to high total IgE levels or body mass. In this study, we sought...

    journal_title:mAbs

    pub_type: 杂志文章

    doi:10.4161/mabs.28394

    authors: Cohen ES,Dobson CL,Käck H,Wang B,Sims DA,Lloyd CO,England E,Rees DG,Guo H,Karagiannis SN,O'Brien S,Persdotter S,Ekdahl H,Butler R,Keyes F,Oakley S,Carlsson M,Briend E,Wilkinson T,Anderson IK,Monk PD,von Wachenfe

    更新日期:2014-05-01 00:00:00