C-terminal lysine variants in fully human monoclonal antibodies: investigation of test methods and possible causes.

Abstract:

:The C-terminal lysine variation is commonly observed in biopharmaceutical monoclonal antibodies. This modification can be important since it is found to be sensitive to the production process. The methods commonly used to probe this charge variation, including IEF, cIEF, ion-exchange chromatography, and LC-MS, were evaluated for their ability to effectively approximate relative percentages of lysine variants. A monoclonal antibody produced in a B cell hybridoma versus a CHO cell transfectoma was examined and it was determined that the relative amount of incorporated C-terminal lysine can vary greatly between these two production schemes. Another case study is shown whereby a different monoclonal antibody is subject to some minor process changes and the extent of lysine variation also exhibits a significant difference. During these studies the different methods for determining the extent of variation were evaluated and it was determined that LC-MS after trypsin digestion provides reproducible relative percentage information and has significant advantages over other methods. The final section of this work investigates the possible origins of this modification and evidence is shown that carboxypeptidase B or another basic carboxypeptidase causes this variation.

journal_name

Biotechnol Bioeng

authors

Dick LW Jr,Qiu D,Mahon D,Adamo M,Cheng KC

doi

10.1002/bit.21855

subject

Has Abstract

pub_date

2008-08-15 00:00:00

pages

1132-43

issue

6

eissn

0006-3592

issn

1097-0290

journal_volume

100

pub_type

杂志文章
  • A reductionist metastasis-on-a-chip platform for in vitro tumor progression modeling and drug screening.

    abstract::Current animal and 2-D cell culture models employed in metastasis research and drug discovery remain poor mimics of human cancer physiology. Here we describe a "metastasis-on-a-chip" system allowing real time tracking of fluorescent colon cancer cells migrating from hydrogel-fabricated gut constructs to downstream liv...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.25950

    authors: Skardal A,Devarasetty M,Forsythe S,Atala A,Soker S

    更新日期:2016-09-01 00:00:00

  • Investigation of particle-functionalized tissue engineering scaffolds using X-ray tomographic microscopy.

    abstract::A low-density, porous chitosan/poly-(dl-lactide-co-glycolide) (PLGA) microparticle composite scaffold was produced by thermally induced phase separation followed by lyophilization, to provide a bicontinuous microstructure potentially suitable for tissue engineering and locally controlled drug release. PLGA particles w...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.21796

    authors: Nygaard JV,Andersen MØ,Howard KA,Foss M,Bünger C,Kjems J,Besenbacher F

    更新日期:2008-07-01 00:00:00

  • Physiologic mechanisms of sequential products synthesis in 1,3-propanediol fed-batch fermentation by Klebsiella pneumoniae.

    abstract::The glycerol fed-batch fermentation by Klebsiella pneumoniae CGMCC 1.6366 exhibited the sequential synthesis of products, including acetate, 1,3-propanediol (1,3-PD), 2,3-butanediol, ethanol, succinate, and lactate. The dominant flux distribution was shifted from acetate formation to 1,3-PD formation in early- exponen...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.21830

    authors: Zheng ZM,Xu YZ,Liu HJ,Guo NN,Cai ZZ,Liu DH

    更新日期:2008-08-01 00:00:00

  • Real-time detection of 13C NMR labeling kinetics in perfused EMT6 mouse mammary tumor cells and betaHC9 mouse insulinomas.

    abstract::A method was developed for obtaining high signal-to-noise 13C NMR spectra of intracellular compounds in metabolically active cultured cells. The method allows TCA cycle labeling kinetics to be determined in real time without significant oxygen transport limitations. Cells were immobilized on the surface of nonporous m...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.20191

    authors: Mancuso A,Beardsley NJ,Wehrli S,Pickup S,Matschinsky FM,Glickson JD

    更新日期:2004-09-30 00:00:00

  • Development of waxy cassava with different Biological and physico-chemical characteristics of starches for industrial applications.

    abstract::The quality of cassava starch, an important trait in cassava breeding programs, determines its applications in various industries. For example, development of waxy (having a low level of amylose) cassava is in demand. Amylose is synthesized by granule-bound starch synthase I (GBSSI) in plants, and therefore, down-regu...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.23120

    authors: Zhao SS,Dufour D,Sánchez T,Ceballos H,Zhang P

    更新日期:2011-08-01 00:00:00

  • Expression of recombinant protein A from the lac promoter in Escherichia coli JM83 is not subject to catabolite repression when grown under specific conditions of continuous culture.

    abstract::Although widely used in experimental and industrial situations, genetically engineered plasmids containing the lac promoter from Escherichia coli are subject to catabolite repression when grown in glucose-containing media. Several methods of overcoming this problem have been investigated by studying the expression of ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260380914

    authors: Warnes A,Stephenson JR,Fooks AR,Melling J,Brown MR

    更新日期:1991-11-01 00:00:00

  • Kinetics of ethanol inhibition in alcohol fermentation.

    abstract::The inhibitory effect of ethanol on yeast growth and fermentation has been studied for the strain Saccharomyces cerevisiae ATCC No. 4126 under anaerobic batch conditions. The results obtained reveal that there is no striking difference between the response of growth and ethanol fermentation. Two kinetic models are als...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260270311

    authors: Luong JH

    更新日期:1985-03-01 00:00:00

  • A multi-site metastasis-on-a-chip microphysiological system for assessing metastatic preference of cancer cells.

    abstract::Metastatic disease remains one of the primary reasons for cancer-related deaths, yet the majority of in vitro cancer models focus on the primary tumor sites. Here, we describe a metastasis-on-a-chip device that houses multiple bioengineered three-dimensional (3D) organoids, established by a 3D photopatterning techniqu...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.26871

    authors: Aleman J,Skardal A

    更新日期:2019-04-01 00:00:00

  • Immobilized phenylalanine hydroxylase through the SH groups.

    abstract::Purified rat liver phenylalanine hydroxylase [L-phenylalanine:tetrahydropteridine:oxygen oxidoreductase (4-hydroxylating), EC 1.14.16.1] was immobilized with activated thiol-Sepharose 4B via disulfide bond formation, which is expected to immobilize the enzyme in its activated form through the SH modification. This imm...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260280422

    authors: Koizumi S,Takeuchi T,Umezawa H,Nagatsu T

    更新日期:1986-04-01 00:00:00

  • Random mutagenesis of global transcription factor cAMP receptor protein for improved osmotolerance.

    abstract::The naturally existing microbial hosts can rarely satisfy industrial requirements, thus there has always been an intense effort in strain engineering to meet the needs of these bioprocesses. Here, in this work, we want to prove the concept that engineering global transcription factor cAMP receptor protein (CRP) of Esc...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.24411

    authors: Zhang H,Chong H,Ching CB,Jiang R

    更新日期:2012-05-01 00:00:00

  • Deregulation of methanol metabolism reverts transcriptional limitations of recombinant Pichia pastoris (Komagataella spp) with multiple expression cassettes under control of the AOX1 promoter.

    abstract::The methanol-regulated alcohol oxidase promoter (PAOX1 ) of Pichia pastoris (syn. Komagataella spp. ) is one of the strongest promoters for heterologous gene expression. Although increasing the gene dosage is a common strategy to improve recombinant protein productivities, P. pastoris strains harboring more than two c...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.26947

    authors: Cámara E,Monforte S,Albiol J,Ferrer P

    更新日期:2019-07-01 00:00:00

  • Synthesis of 3-tert-butylcatechol by an engineered monooxygenase.

    abstract::Recombinant Escherichia coli JM101 was used for the in vivo biocatalytic synthesis of 3-tert-butyl- catechol. The bacterial strain synthesized the laboratory-evolved variant HbpA(T2) of 2-hydroxybiphenyl 3-monooxygenase (HbpA, EC 1.14.13.44) from Pseudomonas azelaica HBP1. The mutant enzyme HbpA(T2) is able to hydroxy...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.10487

    authors: Meyer A,Held M,Schmid A,Kohler HP,Witholt B

    更新日期:2003-03-05 00:00:00

  • Reasons for the apparent difference in the effects of produced and added ethanol on culture viability during rapid fermentation by Saccharomyces cerevisiae.

    abstract::By feeding ethanol at various high rates to low cell density cultures of Saccharomyces cerevisiae it was shown that the sharp fall in viability when ethanol is produced during rapid fermentations is in part a direct consequence of the high rate of change of extracellular ethanol concentration. Nevertheless, the fall i...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260350202

    authors: Dasari G,Worth MA,Connor MA,Pamment NB

    更新日期:1990-01-20 00:00:00

  • Culture of insect cells in helical ribbon impeller bioreactor.

    abstract::An 11-L helical ribbon impeller (HRI) bioreactor was tested for the culture of Spodoptera frugiperda (Sf-9) cells. This impeller and surface baffling ensured homogeneous mixing and high oxygen transfer through surface aeration and surface-induced babble generation. Serum-supplemented and serum-free cultures, using TNM...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260380607

    authors: Kamen AA,Tom RL,Caron AW,Chavarie C,Massie B,Archambault J

    更新日期:1991-09-01 00:00:00

  • Biodegradation of crude oil across a wide range of salinities by an extremely halotolerant bacterial consortium MPD-M, immobilized onto polypropylene fibers.

    abstract::The bacterial consortium MPD-M, isolated from sediment associated with Colombian mangrove roots, was effective in the treatment of hydrocarbons in water with salinities varying from 0 to 180 g L(-1). Where the salinity of the culture medium surpassed 20 g L(-1), its effectiveness increased when the cells were immobili...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.10318

    authors: Díaz MP,Boyd KG,Grigson SJ,Burgess JG

    更新日期:2002-07-20 00:00:00

  • A combinatorial synthesis of tyrphostins via the "directed sorting" method.

    abstract::Using solid-phase organic synthesis, we have prepared a 432-member (18 x 8 x 3) sample library based on the AG 490 "tyrphostin" template. By utilizing 432 reactors each equipped with a unique radiofrequency memory ID tag, the 432 products could be obtained as discrete entities (i.e., not as mixtures) via 18 + 8 + 3, o...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/(sici)1097-0290(199824)61:1<7::aid-bit4>3.

    authors: Shi S,Xiao XY,Czarnik AW

    更新日期:1998-01-01 00:00:00

  • Simulation of oxygen carrier mediated oxygen transport to C3A hepatoma cells housed within a hollow fiber bioreactor.

    abstract::A priori knowledge of the dissolved oxygen (O2) concentration profile within a hepatic hollow fiber (HF) bioreactor is important in developing an effective bioartificial liver assist device (BLAD). O2 provision is limiting within HF bioreactors and we hypothesize that supplementing a hepatic HF bioreactor's circulatin...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.20673

    authors: Sullivan JP,Gordon JE,Palmer AF

    更新日期:2006-02-05 00:00:00

  • Kinetic analysis of bacterial bioluminescence.

    abstract::Bioluminescence from the lux-based bacterial reporter Pseudomonas fluorescens HK44 was experimentally investigated under growth substrate-rich and limiting conditions in batch, continuous stirred tank (CSTR), and turbidostat reactors. A mechanistically based, mathematical model was developed to describe bioluminescenc...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.10475

    authors: Kelly CJ,Hsiung CJ,Lajoie CA

    更新日期:2003-02-05 00:00:00

  • Transformation of 2,4,6-trinitrotoluene (TNT) by immobilized Phanerochaete chrysosporium under fed-batch and continuous TNT feeding conditions.

    abstract::The cometabolic transformation of 2,4,6-trinitrotoluene (TNT) by an immobilized Phanerochaete chrysosporium culture was investigated under different TNT and/or glycerol feeding conditions in a 5-L reactor. In the fed-batch feeding mode, as a result of four spiking events at an average feeding rate of 20 mg TNT L(-1) d...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.1060

    authors: Rho D,Hodgson J,Thiboutot S,Ampleman G,Hawari J

    更新日期:2001-05-20 00:00:00

  • Characterization of extremely thermostable enzymatic breakers (alpha-1,6-galactosidase and beta-1,4-mannanase) from the hyperthermophilic bacterium Thermotoga neapolitana 5068 for hydrolysis of guar gum.

    abstract::An alpha-galactosidase and a beta-mannanase produced by the hyperthermophilic bacterium, Thermotoga neapolitana 5068 (TN5068), separately and together, were evaluated for their ability to hydrolyze guar gum in relation to viscosity reduction of guar-based hydraulic fracturing fluids used in oil and gas well stimulatio...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/(SICI)1097-0290(19961020)52:2<332::AID-BIT

    authors: McCutchen CM,Duffaud GD,Leduc P,Petersen AR,Tayal A,Khan SA,Kelly RM

    更新日期:1996-10-20 00:00:00

  • Diffusivity of oxygen into carriers entrapping whole cells.

    abstract::The effective diffusivity of oxygen, D(e), in Ca-alginate and PVA-SbQ gels was measured using a two-chamber vessel with a membrane between the two chambers. The effect of cell density, C(c), on D(e) in Ca-alginate gels was studied. The effective diffusivity of oxygen decreased with increasing cell density, to C(c) = 1...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260340107

    authors: Sun Y,Furusaki S,Yamauchi A,Ichimura K

    更新日期:1989-06-05 00:00:00

  • Invertase biosynthesis by Saccharomyces carlsbergensis in batch and continuous cultures.

    abstract::The biosynthesis of invertase by Saccharomyces carlsbergensis LAM 1068 was studied in relation to its glucose effect at both unsteady and steady states of growth. Experimental correlations between the dilution rate and invertase specific activity (E/X) in chemostat cultures led to an optimum for the enzyme synthesis a...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260180807

    authors: Toda K

    更新日期:1976-08-01 00:00:00

  • Cellulase activity in commercial lipase preparations.

    abstract::Polymeric membranes are increasingly used as supports for the immobilization of enzymes in bioreactors. One of the more common reactor types employed in lipase-catalyzed hydrolysis of oils, contains modified cellulose as a membrane material. We found that this type of material is readily attacked by cellulase present ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260400714

    authors: Derksen JT,Boswinkel G,Cuperus FP

    更新日期:1992-10-05 00:00:00

  • Interspecific interactions in a methane-utilizing mixed culture.

    abstract::A two-member methane-utilizing mixed culture of bacteria, formed by combining two pure cultures isolated from a naturally occurring methane-utilizing mixed culture, was studied in continuous culture. From the nutritional requirements and substrate ranges of the pure cultures, a mechanism for the interspecific interact...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260221010

    authors: Lamb SC,Garver JC

    更新日期:1980-10-01 00:00:00

  • Escherichia coil growth dynamics: A three-pool biochemically based description.

    abstract::A three-pool growth model of an individual Escherichia coli cell is described herein. The model is based on a previously developed chemically structured complex single cell growth model. The reduction in model complexity and the identification of the essential modes of motion, over the time scale of growth, is achieve...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.260310203

    authors: Joshi A,Palsson BO

    更新日期:1988-02-05 00:00:00

  • Further improvement of phosphite dehydrogenase thermostability by saturation mutagenesis.

    abstract::Phosphite dehydrogenase represents a new enzymatic system for regenerating reduced nicotinamide cofactors for industrial biocatalysis. We previously engineered a variant of phosphite dehydrogenase with relaxed cofactor specificity and significantly increased activity and stability. Here we performed one round of rando...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.21546

    authors: McLachlan MJ,Johannes TW,Zhao H

    更新日期:2008-02-01 00:00:00

  • An actively mixed mini-bioreactor for protein production from suspended animal cells.

    abstract::Biopharmaceutical production would benefit from rapid methods to optimize production of therapeutic proteins by screening host cell line/vector combination, culture media, and operational parameters such as timing of induction. Miniaturized bioreactors are an emerging research area aiming at improving the development ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.21751

    authors: Diao J,Young L,Zhou P,Shuler ML

    更新日期:2008-05-01 00:00:00

  • Integrated single-cell analysis shows Pichia pastoris secretes protein stochastically.

    abstract::The production of heterologous proteins by secretion from cellular hosts is an important determinant for the cost of biotherapeutics. A single-cell analytical method called microengraving was used to examine the heterogeneity in secretion by the methylotrophic yeast Pichia pastoris. We show that constitutive secretion...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.22688

    authors: Love KR,Panagiotou V,Jiang B,Stadheim TA,Love JC

    更新日期:2010-06-01 00:00:00

  • Integrated process design for biocatalytic synthesis by a Leloir Glycosyltransferase: UDP-glucose production with sucrose synthase.

    abstract::Nucleotide sugar-dependent ("Leloir") glycosyltransferases (GTs), represent a new paradigm for the application of biocatalytic glycosylations to the production of fine chemicals. However, it remains to be shown that GT processes meet the high efficiency targets of industrial biotransformations. We demonstrate in this ...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.26204

    authors: Schmölzer K,Lemmerer M,Gutmann A,Nidetzky B

    更新日期:2017-04-01 00:00:00

  • Production of cell culture (MDCK) derived live attenuated influenza vaccine (LAIV) in a fully disposable platform process.

    abstract::The majority of influenza vaccines are manufactured using embryonated hens' eggs. The potential occurrence of a pandemic outbreak of avian influenza might reduce or even eliminate the supply of eggs, leaving the human population at risk. Also, the egg-based production technology is intrinsically cumbersome and not eas...

    journal_title:Biotechnology and bioengineering

    pub_type: 杂志文章

    doi:10.1002/bit.22753

    authors: George M,Farooq M,Dang T,Cortes B,Liu J,Maranga L

    更新日期:2010-08-15 00:00:00