Phenolics removal from transgenic <i>Lemna minor</i> extracts expressing mAb and impact on mAb production cost

Barros, G. O. F.; Woodard, Susan L.; Nikolov, Z̆ivko L.

doi:10.1002/btpr.543

Cited by 21 publications

(10 citation statements)

References 26 publications

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“…In principle, most plantibodies are expressed in tobacco ( N. tabacum or N. benthamiana ), but there are also production systems in Lemna minor (duckweed) (209–211), rice cell culture (212), Arabidopsis thaliana seeds (213, 214), Medicago sativa (alfalfa) (215), lettuce (216), and maize (217). HIV-1 neutralizing antibody 2G12 was expressed in the endosperm of maize and showed similar or even better neutralizing properties as its CHO-derived counterpart (218).…”

Section: Transgenic Organismsmentioning

confidence: 99%

Expression of Recombinant Antibodies

Frenzel¹,

Hust²,

Schirrmann³

2013

Front. Immunol.

279

220

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Recombinant antibodies are highly specific detection probes in research, diagnostics, and have emerged over the last two decades as the fastest growing class of therapeutic proteins. Antibody generation has been dramatically accelerated by in vitro selection systems, particularly phage display. An increasing variety of recombinant production systems have been developed, ranging from Gram-negative and positive bacteria, yeasts and filamentous fungi, insect cell lines, mammalian cells to transgenic plants and animals. Currently, almost all therapeutic antibodies are still produced in mammalian cell lines in order to reduce the risk of immunogenicity due to altered, non-human glycosylation patterns. However, recent developments of glycosylation-engineered yeast, insect cell lines, and transgenic plants are promising to obtain antibodies with “human-like” post-translational modifications. Furthermore, smaller antibody fragments including bispecific antibodies without any glycosylation are successfully produced in bacteria and have advanced to clinical testing. The first therapeutic antibody products from a non-mammalian source can be expected in coming next years. In this review, we focus on current antibody production systems including their usability for different applications.

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Section: Transgenic Organismsmentioning

confidence: 99%

Expression of Recombinant Antibodies

Frenzel¹,

Hust²,

Schirrmann³

2013

Front. Immunol.

279

220

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“…We used the MagnICON Δ87GAD65mut because of its superior yield and solubility and MagnICON full‐length GAD65mut for comparison. We measured extract turbidity, TSP and the content of phenolic compounds .…”

Section: Resultsmentioning

confidence: 99%

Enhanced GAD65 production in plants using the MagnICON transient expression system: Optimization of upstream production and downstream processing

Merlin

Gecchele

Arcalís

et al. 2016

Biotechnology Journal

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Plants have emerged as competitive production platforms for pharmaceutical proteins that are required in large quantities. One example is the 65-kDa isoform of human glutamic acid decarboxylase (GAD65), a major autoimmune diabetes autoantigen that has been developed as a vaccine candidate for the primary prevention of diabetes. The expression of GAD65 in plants has been optimized but large-scale purification is hampered by its tendency to associate with membranes. We investigated the potential for large-scale downstream processing by evaluating different combinations of plant-based expression systems and engineered forms of GAD65 in terms of yield, subcellular localization and solubility in detergent-free buffer. We found that a modified version of GAD65 lacking the first 87 amino acids accumulates to high levels in the cytosol and can be extracted in detergent-free buffer. The highest yields of this variant protein were achieved using the MagnICON transient expression system. This combination of truncated GAD65 and the MagnICON system dramatically boosts the production of the recombinant protein and helps to optimize downstream processing for the establishment of a sustainable plant-based production platform for an autoimmune diabetes vaccine candidate.

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“…As most so-called plantibodies are expressed in tobacco plants, several other species have been used as expression system such as Arabidopsis thaliana [256], duckweed [257,258], lettuce [259], maize [260], Medicago sativa [261], and rice cell culture [262].…”

Section: Plantsmentioning

confidence: 99%

Emerging Alternative Production Systems

Sommer

Laux

Frenzel

et al. 2014

Handbook of Therapeutic Antibodies

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The market of recombinant protein pharmaceuticals is growing rapidly. Among those, recombinant antibodies are the fastest growing group and a large number of new antibody products are in clinical and preclinical development. Since most conventional cost-effective microbial expression systems are not yet applicable for the expression of fully functional immunoglobulin G (IgG) antibodies, almost all of the currently marketed therapeutic recombinant antibody products are produced from animal cell culture processes that are associated with relative high operating expenses. Thus, to make broad use of recombinant antibody therapeutics more affordable, alternative production systems with improved efficiency and reduced operating expenses are highly desirable.IgG currently is the dominant antibody format for therapeutics. As a heterotetrameric molecule with numerous disulfide bonds and several glycosylation sites IgG makes itself a challenging candidate for heterologous expression, especially in prokaryotes. Glycosylation of the fragment crystallizable (Fc) region has been shown to be essential for mediating effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) [1]. Furthermore, the glycosylation pattern strongly influences the efficiency of the induction of effector functions by antibodies [2, 3]. For applications that depend on the immunological effector functions of the antibody therefore eukaryotic production systems with suitable glycosylation capabilities are necessary.Antibody fragments such as single chain fragment variable (scFv) and fragment antigen binding (Fab) are less complex and the antigen-binding activity is independent of glycosylation, which makes them qualified for prokaryotic expression systems. Certainly, these fragments are incapable of mediating immunological effector functions but for numerous applications this is not necessary or even unwanted.Besides the two traditional expression systems, Escherichia coli for antibody fragments and Chinese hamster ovarian (CHO) or myeloma cells for IgG antibodies, numerous alternative expression systems are currently under development.

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Phenolics removal from transgenic Lemna minor extracts expressing mAb and impact on mAb production cost

Cited by 21 publications

References 26 publications

Expression of Recombinant Antibodies

Expression of Recombinant Antibodies

Enhanced GAD65 production in plants using the MagnICON transient expression system: Optimization of upstream production and downstream processing

Emerging Alternative Production Systems

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