当前位置： SCI文献检索 > PROTEIN EXPRESSION AND PURIFICATION期刊下所有文献 > Recombinant production of a VL single domain antibody in Escherichia coli and analysis of its interaction with peptostreptococcal protein L.

Recombinant production of a VL single domain antibody in Escherichia coli and analysis of its interaction with peptostreptococcal protein L.

Abstract：

:A kappa-light chain from a Fab expression system was truncated by the insertion of a stop codon in the gene sequence to produce a variable light (VL) single domain antibody (dAb). Here, we describe the expression of dAb in the periplasm of Escherichia coli through fermentation in a defined media. Immunoglobulin binding domains from peptostreptococcal protein L (PpL) have been shown to bind specifically to kappa-light chains. We have produced recombinant PpL, at high yield, and this was used to custom-produce PpL-Sepharose affinity columns. Here, we show that the affinity purification of VL dAb by this method is simple and efficient with no apparent loss in protein at any stage. The truncated dAb protein product was confirmed by electrospray mass spectrometry and N-terminal sequencing. When analyzed by SDS-PAGE it was shown to be over 95% pure and produced at yields of 35-65 mg/L of culture medium. The dAb protein produced was shown by NMR and CD to be a folded beta-sheet domain. This domain is bound by PpL with a Kd of approximately 50 nM as determined by stopped-flow fluorimetry.

journal_name

Protein Expr Purif

journal_title

Protein expression and purification

authors

Cossins AJ,Harrison S,Popplewell AG,Gore MG

doi

10.1016/j.pep.2006.07.013

subject

Has Abstract

pub_date

2007-02-01 00:00:00

pages

253-9

issue

2

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(06)00226-9

journal_volume

51

pub_type

杂志文章

相关文献

PROTEIN EXPRESSION AND PURIFICATION文献大全
  • Heat stability of Proteobacterial PII protein facilitate purification using a single chromatography step.

    abstract：:The P(II) proteins comprise a family of widely distributed signal transduction proteins that integrate the signals of cellular nitrogen, carbon and energy status, and then regulate, by protein-protein interaction, the activity of a variety of target proteins including enzymes, transcriptional regulators and membrane t...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2011.09.008

    authors： Moure VR,Razzera G,Araújo LM,Oliveira MAS,Gerhardt ECM,Müller-Santos M,Almeida F,Pedrosa FO,Valente AP,Souza EM,Huergo LF

    更新日期：2012-01-01 00:00:00

  • Generation and evaluation of mammalian secreted and membrane protein expression libraries for high-throughput target discovery.

    abstract：:Expressed protein libraries are becoming a critical tool for new target discovery in the pharmaceutical industry. In order to get the most meaningful and comprehensive results from protein library screens, it is essential to have library proteins in their native conformation with proper post-translation modifications....

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2011.05.011

    authors： Panavas T,Lu J,Liu X,Winkis AM,Powers G,Naso MF,Amegadzie B

    更新日期：2011-09-01 00:00:00

  • Affinity purification of ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit epsilon N-methyltransferase.

    abstract：:Ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit epsilon N-methyltransferase (Protein methylase III, Rubisco LSMT, EC 2.1.1.43) catalyzes methylation of the epsilon-amino group of Lys-14 in the large subunit of Rubisco. In this paper, an affinity purification procedure for pea (Pisum sativum L. cv Laxton'...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1995.1070

    authors： Wang P,Royer M,Houtz RL

    更新日期：1995-08-01 00:00:00

  • Rapid high-yield purification and liposome reconstitution of polyhistidine-tagged sensory rhodopsin I.

    abstract：:We have used Ni(2+)-affinity chromatography as a rapid and efficient method to purify a sensory rhodopsin I (SR-I) derivative containing six consecutive histidine residues at its C-terminus (His-tagged SR-I). The protein was expressed in Halobacterium salinarium by integrating the corresponding gene at the chromosomal...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1995.0009

    authors： Krebs MP,Spudich EN,Spudich JL

    更新日期：1995-12-01 00:00:00

  • Production of correctly folded Fab antibody fragment in the cytoplasm of Escherichia coli trxB gor mutants via the coexpression of molecular chaperones.

    abstract：:Disulfide bonds are normally formed after a polypeptide has been exported from the reducing environment of the cytoplasm into a more oxidizing compartment, such as the bacterial periplasm. Recently, we showed that in Escherichia coli trxB gor mutants, in which the reduction of thioredoxin and glutathione is impaired, ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2001.1520

    authors： Levy R,Weiss R,Chen G,Iverson BL,Georgiou G

    更新日期：2001-11-01 00:00:00

  • Cholera toxin B subunit-domain III of dengue virus envelope glycoprotein E fusion protein production in transgenic plants.

    abstract：:Envelope glycoprotein E of the dengue virus, which plays a crucial role in its entry into host cells, has an immunogenic domain III (EIII, amino acids 297-394), which is capable of inducing neutralizing antibodies. However, mice immunized with EIII protein without adjuvant elicited low immune responses. To improve low...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2010.07.013

    authors： Kim TG,Kim MY,Yang MS

    更新日期：2010-12-01 00:00:00

  • Expression, purification and characterization of heterotrimeric forms of sTRAIL using a polycistronic expression vector.

    abstract：:Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), which is capable of selectively inducing apoptosis of cancer cells, is a potential targeted drug for cancer therapy. The TRAIL protein induces apoptosis only in trimeric form. However, the recombinant soluble TRAIL (sTRAIL) trimer has low stability and a...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2015.08.004

    authors： Wang B,Wang Z,Yan J,Wang L,Wang Z,Wu J,Zhang H,Wu H,Kong W,Yu B,Yu X

    更新日期：2015-11-01 00:00:00

  • Expression, purification, and characterization of deglycosylated human pro-prostate-specific antigen.

    abstract：:Wild-type and deglycosylated forms of human prostate-specific antigen were expressed in Chinese hamster ovary (CHO) cells as zymogens. ProPSA was collected from conditioned medium and purified using a single cation-exchange chromatographic step for the deglycosylated form and cation-exchange followed by gel filtration...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2000.1342

    authors： Bowman KK,Clark J,Yu L,Mortara K,Radika K,Wang J,Zhan H

    更新日期：2000-12-01 00:00:00

  • Expression of the recombinant antibacterial peptide sarcotoxin IA in Escherichia coli cells.

    abstract：:Sarcotoxin IA is an antibacterial peptide that is secreted by a meat-fly Sarcophaga peregrina larva in response to a hypodermic injury or bacterial infection. This peptide is highly toxic against a broad spectrum of both Gram-positive and Gram-negative bacteria and lethal to microbes even at nanomolar concentrations. ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/s1046-5928(02)00697-6

    authors： Skosyrev VS,Kulesskiy EA,Yakhnin AV,Temirov YV,Vinokurov LM

    更新日期：2003-04-01 00:00:00

  • Expression in Trichoderma reesei and characterisation of a thermostable family 3 beta-glucosidase from the moderately thermophilic fungus Talaromyces emersonii.

    abstract：:The gene encoding a thermostable beta-glucosidase (cel3a) was isolated from the thermophilic fungus Talalaromyces emersonii by degenerate PCR and expressed in the filamentous fungus Trichoderma reesei. The cel3a gene encodes an 857 amino acid long protein with a calculated molecular weight of 90.59 kDa. Tal. emersonii...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2004.08.006

    authors： Murray P,Aro N,Collins C,Grassick A,Penttilä M,Saloheimo M,Tuohy M

    更新日期：2004-12-01 00:00:00

  • Intein-mediated expression, purification, and characterization of thymosin α1-thymopentin fusion peptide in Escherichia coli.

    abstract：:Thymosin α1-thymopentin (Tα1-TP5) fusion peptide has been proved to be an immune regulator based on its higher immunoregulatory activity than Tα1 and TP5. To obtain Tα1-TP5 more effectively and economically, Tα1-TP5 was genetically fused to a self-cleaving intein-chitin binding domain tag for purification via chitin b...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2012.04.013

    authors： Li J,Zheng L,Li P,Wang F

    更新日期：2012-07-01 00:00:00

  • Affinity purification of recombinant human cytochrome P450s 3A4 and 1A2 using mixed micelle systems.

    abstract：:Recombinant cytochrome P450 (CYP or P450) enzymes are useful for drug metabolism research and thereby many expression and purification systems have been developed. Here, we provide a method for the purification of human P450s 3A4 and 1A2 expressed in Escherichia coli using mixed micelles containing anionic phospholipi...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2014.05.010

    authors： Ahn T,Bae CS,Yun CH

    更新日期：2014-09-01 00:00:00

  • Heterologous expression of Class IIb bacteriocin Plantaricin JK in Lactococcus Lactis.

    abstract：:Plantaricin JK (PlnJK) is a Class IIb LAB bacteriocin that includes two peptides; i.e., PlnJ and PlnK, which can synergistically halt many types of gram-positive bacteria, including food spoilage organisms. Purification of these peptides from natural lactic acid bacteria is difficult therefore, their application remai...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2019.02.013

    authors： Xu Y,Yang L,Li P,Gu Q

    更新日期：2019-07-01 00:00:00

  • High-level bacterial expression of human glutathione transferase P1-1 encoded by semisynthetic DNA.

    abstract：:A cDNA clone, lambda GTHP1del, encoding glutathione transferase (GST) P1-1, was isolated from a human K562 erythroleukemia cell line cDNA library. The coding sequence was lacking the codons for the N-terminal 34 amino acids. A DNA segment was designed in order to obtain the missing portion and a structure representing...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1995.1034

    authors： Kolm RH,Stenberg G,Widersten M,Mannervik B

    更新日期：1995-06-01 00:00:00

  • Expression and purification of the alpha-subunit of eukaryotic initiation factor eIF2: use as a kinase substrate.

    abstract：:The alpha-subunit of eukaryotic initiation factor eIF2 (eIF2alpha) plays an important role in the regulation of mRNA translation through modulation of the interaction of eIF2 and a second initiation factor, eIF2B. The interaction of the two proteins is regulated in vivo by phosphorylation of eIF2alpha at Ser51. In the...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1998.0863

    authors： Kimball SR,Horetsky RL,Jagus R,Jefferson LS

    更新日期：1998-04-01 00:00:00

  • High-level synthesis of recombinant murine endostatin in Chinese hamster ovary cells.

    abstract：:Endostatin, a carboxy-terminal fragment of collagen XVIII, has been shown to act as an anti-angiogenic agent that specifically inhibits proliferation of endothelial cells and growth of various primary tumors. Here, we describe the expression by Chinese hamster ovary (CHO) cells of murine endostatin and of a tagged-fus...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2004.01.003

    authors： Chura-Chambi RM,Tornieri PH,Spencer PJ,Nascimento PA,Mathor MB,Morganti L

    更新日期：2004-05-01 00:00:00

  • Streamlined procedure for the production of normal and altered versions of recombinant human proinsulin.

    abstract：:A method for the simplified, reproducible production of both normal and altered versions of human proinsulin has been developed. A polyhistidine/proinsulin fusion protein was expressed using a prokaryotic expression system and partially purified by affinity chromatography. Disulfide bonds within the polypeptide were f...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1998.1024

    authors： Mackin RB

    更新日期：1999-04-01 00:00:00

  • Effect of K225 residue to the catalytic efficiency of Kex2 protease.

    abstract：:The gene encoding S. cerevisiae Kex2 protease derivative Kex2-667 (encoding the N-terminal 20th to 667th amino acid residues of Kex2 protease, containing the propeptide, catalytic domain, P domain and Ser/Thr enrichment region) and its 225th amino acid residue mutant K225L were overexpressed in Pichia pastoris. Protea...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2020.105725

    authors： Yang F,Liu L,Liu Y,Li S

    更新日期：2020-12-01 00:00:00

  • Overexpression, purification, and characterization of a thermostable chitinase (Chi40) from Streptomyces thermoviolaceus OPC-520.

    abstract：:A new procedure for the large-scale purification of the recombinant thermostable chitinase (Chi40) cloned from Streptomyces thermoviolaceus in various expression vectors in Escherichia coli is described. Chi40 was overproduced in the cytosolic and secreted forms. The cytosolic form (Chi40c) was highly overproduced and...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2001.1490

    authors： Christodoulou E,Duffner F,Vorgias CE

    更新日期：2001-10-01 00:00:00

  • Peptidyl-prolyl isomerase and the biological activities of recombinant protein cyclophilin from Pyropia yezoensis (PyCyp).

    abstract：:Cyclophilins are highly conserved proteins associated with peptidyl-prolyl cis-trans isomerase activity (PPIase). The present study was designed to analyze the biological activity of recombinant cyclophilin from the marine red algae Pyropia yezoensis (PyCyp). The cyclophilin gene from P. yezoensis was cloned into the ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2020.105636

    authors： Ulagesan S,Choi JW,Nam TJ,Choi YH

    更新日期：2020-08-01 00:00:00

  • The Nano-tag, a streptavidin-binding peptide for the purification and detection of recombinant proteins.

    abstract：:We present a new streptavidin-binding peptide for both the purification and the detection of recombinant proteins. The peptide possesses nanomolar-affinity for streptavidin and therefore was termed Nano-tag. The Nano-tag(15) is 15 amino acids long and binds to streptavidin with a dissociation constant of 4 nM and the ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2003.08.014

    authors： Lamla T,Erdmann VA

    更新日期：2004-01-01 00:00:00

  • Expression, purification, and initial structural characterization of nonstructural protein 2B, an integral membrane protein of Dengue-2 virus, in detergent micelles.

    abstract：:Dengue virus causes serious diseases affecting people in tropical and sub-tropical regions. The nonstructural (NS) protein 2B is an integral membrane protein and important for the regulation of viral protease NS3, which is significant for virus replication. The NS2B-NS3 complex is an important drug target for treating...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2011.08.008

    authors： Huang Q,Chen AS,Li Q,Kang C

    更新日期：2011-12-01 00:00:00

  • Engineering S-protein fragments of bovine ribonuclease A for targeted drug delivery.

    abstract：:High affinity interaction between S-protein and S-peptide fragments of bovine pancreatic RNase A has been recently used for construction of molecular vehicles for targeted drug delivery. The vehicle is assembled as a complex of drug carrier conjugated S-protein with S-peptide-tagged targeting protein. To avoid random ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/s1046-5928(02)00546-6

    authors： Backer MV,Gaynutdinov TI,Aloise R,Przekop K,Backer JM

    更新日期：2002-12-01 00:00:00

  • Efficient recovery of the functional IP10-scFv fusion protein from inclusion bodies with an on-column refolding system.

    abstract：:A functional IP10-scFv fusion protein retaining the antibody specificity for acidic isoferritin and chemokine function was produced at high level in Esherichia coli (E. coli). IP10-scFv gene from the recombinant plasmid pc3IP104c9 was subcloned into pET28a fused to N-terminal His-tag sequence in frame and overexpresse...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2005.05.016

    authors： Guo JQ,Li QM,Zhou JY,Zhang GP,Yang YY,Xing GX,Zhao D,You SY,Zhang CY

    更新日期：2006-01-01 00:00:00

  • Efficient expression of SRK intracellular domain by a modeling-based protein engineering.

    abstract：:S-locus protein kinase (SRK) is a receptor kinase that plays a critical role in self-recognition in the Brassicaceae self-incompatibility (SI) response. SRK is activated by binding of its ligand S-locus protein 11 (SP11) and subsequently induced phosphorylation of the intracellular kinase domain. However, a detailed a...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2015.09.020

    authors： Murase K,Hirano Y,Takayama S,Hakoshima T

    更新日期：2017-03-01 00:00:00

  • Expression, purification, and characterization of an unstable lysozyme mutant in Pichia pastoris.

    abstract：:To investigate the expression and purification of an unstable heterologous protein in Pichia pastoris, the cDNA of H5-lysozyme, a hen egg lysozyme mutant with a hydrophobic pentapeptide (Phe-Phe-Val-Ala-Pro) fused to the carboxyl terminus, was integrated into the genome of P. pastoris. It was found that medium composi...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/s1046-5928(02)00642-3

    authors： Liu ST,Saito A,Azakami H,Kato A

    更新日期：2003-02-01 00:00:00

  • Solubility of the catalytic domains of Botulinum neurotoxin serotype E subtypes.

    abstract：:The Clostridium botulinum neurotoxins (BoNTs) are the most potent protein toxins known to humans. There are seven serotypes of the BoNTs (A-G), among which serotypes A, B, E and F are known to cause natural human intoxication. To date, eleven subtypes of LC/E, termed E1∼E11, have been identified. The LCs of BoNT/E wer...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2015.10.003

    authors： Chen S,Barbieri JT

    更新日期：2016-02-01 00:00:00

  • Cloning, expression and characterization of the recombinant Yersinia pseudotuberculosis L-asparaginase.

    abstract：:We have cloned ansB (YPTB1411) gene from Yersinia pseudotuberculosis Q66CJ2 and constructed stable inducible expression system that overproduce L-asparaginase from Y. pseudotuberculosis (YpA) in Escherichiacoli BL21 (DE3) cells. For purification of YpA we used Q-Sepharose and DEAE-Toyopearl column chromatography. We e...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2011.12.005

    authors： Pokrovskaya MV,Aleksandrova SS,Pokrovsky VS,Omeljanjuk NM,Borisova AA,Anisimova NY,Sokolov NN

    更新日期：2012-03-01 00:00:00

  • Cloning, expression and functional characterization of two sesquiterpene synthase genes from moso bamboo (Phyllostachys edulis).

    abstract：:The purpose of this work was to characterize the functions of two sesquiterpene synthase genes from moso bamboo (Phyllostachys edulis). Two novel sesquiterpene synthase genes, belonging to the Tpsa subfamily, were isolated from moso bamboo. MoTPS2 was 1641 bp in length and encoded a protein of 63 kDa, whereas MoTPS6 w...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2015.11.019

    authors： Chen X,Wang Y,Sun J,Wang J,Xun H,Tang F

    更新日期：2016-04-01 00:00:00

  • Purification and characterization of a long-acting ciliary neurotrophic factor via genetically fused with an albumin-binding domain.

    abstract：:Ciliary neurotrophic factor (CNTF) is a promising candidate for the treatment of neurodegenerative or metabolic diseases, but suffers rapid clearance in body. Herein we constructed a new long-acting recombinant human CNTF (rhCNTF) by genetic fusion with an albumin-binding domain (ABD) through a flexible peptide linker...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2017.07.006

    authors： Xu L,Zhang C,Liu L,Zhang Y,Wang Q,Wang J,Liu Y,Su Z

    更新日期：2017-11-01 00:00:00