An optimized fermentation process for high-level production of a single-chain Fv antibody fragment in Pichia pastoris

Damasceno, Leonardo M.; Pla, Itzcoatl; Chang, Hyun-Joo; Cohen, Lawrence E.; Ritter, Gerd; Old, Lloyd J.; Batt, Carl A.

doi:10.1016/j.pep.2004.03.019

Cited by 121 publications

(91 citation statements)

References 27 publications

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“…In the applied sciences, biosensors increasingly contribute in clinical [886][887][888][889][890][891][892][893][894][895][896][897][898][899][900][901][902][903] and food/veterinary [904][905][906][907][908][909][910][911][912][913] settings. This year's less-traditional uses of the biosensor include coupling Biacore with mass spectrometry [914][915][916], examining intricate systems such as fibril, polymer and nanotube construction and elongation, monitoring stages of bioprocessing, and characterizing constituents of venom, lysates and other crude analytes [917][918][919][920][921][922][923][924][925][926][927][928][929][930][931].…”

Section: Instrumentsmentioning

confidence: 99%

Survey of the year 2004 commercial optical biosensor literature

Rich

Myszka

2005

J of Molecular Recognition

115

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The year 2004 represents a milestone for the biosensor research community: in this year, over 1000 articles were published describing experiments performed using commercially available systems. The 1038 papers we found represent a $ 10% increase over the past year and demonstrate that the implementation of biosensors continues to expand at a healthy pace. We evaluated the data presented in each paper and compiled a 'top 10' list. These 10 articles, which we recommend every biosensor user reads, describe wellperformed kinetic, equilibrium and qualitative/screening studies, provide comparisons between binding parameters obtained from different biosensor users, as well as from biosensor-and solution-based interaction analyses, and summarize the cutting-edge applications of the technology. We also re-iterate some of the experimental pitfalls that lead to sub-optimal data and over-interpreted results. We are hopeful that the biosensor community, by applying the hints we outline, will obtain data on a par with that presented in the 10 spotlighted articles. This will ensure that the scientific community at large can be confident in the data we report from optical biosensors.

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Section: Instrumentsmentioning

confidence: 99%

Survey of the year 2004 commercial optical biosensor literature

Rich

Myszka

2005

J of Molecular Recognition

115

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“…These alternative transgenic expression systems could reduce the cost of large-scale antibody production. However, the prolonged construction of transgenics remains a major disadvantage in terms of market demands.The production of antibodies and antibody fragments has been studied by using various microorganism expression systems, including Escherichia coli, fungus, and yeast (7,13,37,41). Since microorganisms are known to possess certain advantages as hosts (e.g., rapid growth rate, high heterologous protein productivity, and inexpensive cultivation costs), they have great potential to overcome some of the current hindrances to antibody production.…”

mentioning

confidence: 99%

Efficient Antibody Production upon Suppression of O Mannosylation in the Yeast Ogataea minuta

Kuroda

Kobayashi

Kitagawa

et al. 2008

Appl Environ Microbiol

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When antibodies were expressed in the methylotrophic yeast Ogataea minuta, we found that abnormal O mannosylation occurred in the secreted antibody. Yeast-specific O mannosylation is initiated by the addition of mannose at serine (Ser) or threonine (Thr) residues in the endoplasmic reticulum via protein O mannosyltransferase (Pmt) activity. To suppress the addition of O-linked sugar chains on antibodies, we examined the possibility of inhibiting Pmt activity by the addition of a Pmt inhibitor during cultivation. The Pmt inhibitor was found to partially suppress the O mannosylation on the antibodies. Surprisingly, the suppression of O mannosylation was associated with an increased amount of assembled antibody (H2L2) and enhanced the antigen-binding activity of the secreted antibody. In this study, we demonstrated the expression of human antibody in O. minuta and elucidated the relationship between O mannosylation and antibody production in yeast.Antibodies for pharmaceutical applications have recently received a great deal of attention due to their specific antigenbinding activities, antibody-dependent cellular cytotoxicity, and other useful characteristics. Indeed, sales of antibodies for pharmaceuticals have grown rapidly over the past decade and are expected to exceed 30 billion U.S. dollars by 2010 (1). A large volume of antibodies is required for pharmaceuticals, as antibodies are primarily marketed for chronic conditions. Antibodies possess relatively low potency, which results in the need for accumulating doses over time (10). Although therapeutic antibodies have great potential value, they have also been viewed with some skepticism. Antibodies are expensive to produce, in part because they are commonly manufactured by using batch/fed-batch cultures of mammalian cells. The increasing demand to reduce the cost of antibody production has promoted research into the development of an antibody expression system that is more practical than expression with mammalian cells.Development of transgenic plants and animals as alternative hosts is therefore a promising field of study. Monoclonal antibodies have, thus far, been successfully produced from a number of sources, including plants, the milk of transgenic goats, the eggs of transgenic chickens, etc. (4, 12, 32). Moreover, antibodies derived from certain newly developed transgenic systems share physical characteristics that are similar to those of antibodies from mammalian cells such as Chinese hamster ovary (CHO) cells while exhibiting higher antibody-dependent cellular cytotoxicity activity due to the absence of fucose residues in N-linked sugar chains (6, 45). These alternative transgenic expression systems could reduce the cost of large-scale antibody production. However, the prolonged construction of transgenics remains a major disadvantage in terms of market demands.The production of antibodies and antibody fragments has been studied by using various microorganism expression systems, including Escherichia coli, fungus, and yeast (7,13,37,41). Since microorg...

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“…hydrogen, ethanol) 12,13 , pharmaceuticals (e.g. antibiotics, diagnostic markers) 14,15 , and industrial chemicals (e.g. organic acids, biopolymers) [16][17][18] .…”

Section: Statement Of Part Of the Thesis Submitted To Qualify For Thementioning

confidence: 99%

Metabolic mechanisms and regulation of mixed culture fermentation

Hoelzle¹

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Fermentation of waste streams is an emerging method for production of biofuels and commodity chemicals, and mixed-culture fermentation allows for robustness and flexibility in the range of waste streams that can be processed, and in varying product mix from the fermenter. However, mechanistic models of mixed culture fermentation are limited, with mixed culture fermentation generally resulting in acetate, butyrate, and ethanol, with even these production modes are not clearly understood. Results from pure-culture fermentation, as well as those from methanogenic reactors indicate the possibility to produce high-value lactate and propionate if the cellular mechanisms which control the expression of these production modes can be harnessed and understood. Lactate and propionate are key chemicals in production of bioplastics, pesticides, preservatives, and food additives, among many other uses. Their production by mixed cultures has been observed at high substrate concentrations (shock loads), and inconsistently at high pH. They are produced via metabolic pathways which branch off of the primary energy-generation pathways. In general, they are thought to be produced by microbes as a way to redirect excess carbon flow in transient states, and are also thought to be produced as an electron sink. In this work, both of these possible production influences are explored for steady state production of lactate and propionate.By increasing substrate from limiting to non-limiting quantities, and thereby increasing the carbon load on a fermentation system, lactate production was successfully achieved and maintained during steady state only when substrate was in excess. Substrate consumption at this point was 14.1±0.8 gCOD·L -1 , and lactate accounted for 25±1% of the total product COD. Upon return to substrate-limiting conditions, the process returned to butyrate production. This demonstrates the reversibility of this production mode. Taxonomic analysis via Illumina pyrosequencing of the 16S rRNA gene revealed that the microbial community experienced a shift from Megasphaera-dominant during butyrate production to Ethanoligenens-dominant during lactate production. Taxonomy was also regained upon return to substrate-limiting conditions. However, taxonomy alone could not explain the product shift because Ethanoligenens produces little to no lactate in cultured references. A mechanistic description of the product shift therefore required molecular analysis of the metabolic mechanisms, and this was achieved through metaproteomics via SWATH-MS.The evidence suggests that at high carbon loads, toxic methylglyoxal is formed in many microbe groups, including Megasphaera, due to limited processing capacity of the ii dihydroxyacetone-P intermediate of glycolysis. The methylglyoxal is converted into lactate for disposal. This detoxification need is abated upon return to substrate-limited conditions. The impact of redox stress was examined using cathodic electro-fermentation with soluble mediator chemicals ranging from slightly negati...

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An optimized fermentation process for high-level production of a single-chain Fv antibody fragment in Pichia pastoris

Cited by 121 publications

References 27 publications

Survey of the year 2004 commercial optical biosensor literature

Survey of the year 2004 commercial optical biosensor literature

Efficient Antibody Production upon Suppression of O Mannosylation in the Yeast Ogataea minuta

Metabolic mechanisms and regulation of mixed culture fermentation

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