scholarly journals cDNA Cloning of a Bovine Insulin-like growth factor-1 from Egyptian Buffalos and Expression of its Recombinant Protein in Escherichia coli

2020 ◽  
Vol 72 (2) ◽  
pp. 523-534
Author(s):  
Mourad A.M. Aboul-Soud
Keyword(s):  
Growth Factor ◽  
Mammalian Cells ◽  
Large Scale ◽  
Recombinant Dna ◽  
Bovine Insulin ◽  
Scale Production ◽  
Insulin Like Growth Factor ◽  
Cdna Insert ◽  
Hela Cell Lines ◽  
Clinically Significant

ABSTRACT Insulin-like growth factor-1 (IGF-1) is regarded as a crucial clinically significant therapeutic agent against several pathological conditions. Recently, recombinant DNA (rDNA) technology has enabled the production of many drugs of rDNA-origin including IGF-1. Securing a readily available supply of IGF-1 is invaluable to clinical research and biotechnological domains. In this work, the cloning of a full-length bovine IGF-1 cDNA and the successful expression of its cognate recombinant IGF-1 protein is reported. Single-strand cDNA was prepared from liver tissues, through the specific reverse transcription (RT) of IGF-1 mRNA. Subsequently, a PCR amplicon of ~543bp was successfully amplified. Recombinant pTARGET™ vector harboring IGF-1 insert was successfully cloned into competent E. coli JM109 cells. SDS-PAGE analysis revealed that the recombinant IGF-1 has been expressed at the expected size of 7.6kDa. The outcome provides a robust basis for transecting the recombinant pTARGETTM vector, harboring the IGF-1 cDNA insert, into mammalian cells. Optimal initial glucose concentration was found to be 10g/l with corresponding protein concentration of 6.2g/l. The proliferative biological activity crude recombinant IGF-1 protein was verified on HeLa cell lines. This is envisaged to facilitate large-scale production of recombinant IGF-1 protein, thereby enabling thorough investigation of its clinical and pharmaceutical effects.

scholarly journals Transgenic goats producing an improved version of cetuximab in milk

2020 ◽  
Author(s):  
Götz Laible ◽  
Sally Cole ◽  
Brigid Brophy ◽  
Paul Maclean ◽  
Li How Chen ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Large Scale ◽  
Growth Factor Receptor ◽  
Cost Effective ◽  
Scale Production ◽  
Cancer Treatments ◽  
Large Scale Production ◽  
Immunogenic Epitope ◽  
Dependent Cell ◽  
Transgenic Goats

ABSTRACTTherapeutic monoclonal antibodies (mAbs) represent one of the most important classes of pharmaceutical proteins to treat human diseases. Most are produced in cultured mammalian cells which is expensive, limiting their availability. Goats, striking a good balance between a relatively short generation time and copious milk yield, present an alternative platform for the cost-effective, flexible, large-scale production of therapeutic mAbs. Here, we focused on cetuximab, a mAb against epidermal growth factor receptor, that is commercially produced under the brand name Erbitux and approved for anti-cancer treatments. We generated several transgenic goat lines that produce cetuximab in their milk. Two lines were selected for detailed characterization. Both showed stable genotypes and cetuximab production levels of up to 10g/L. The mAb could be readily purified and showed improved characteristics compared to Erbitux. The goat-produced cetuximab (gCetuximab) lacked a highly immunogenic epitope that is part of Erbitux. Moreover, it showed enhanced binding to CD16 and increased antibody-dependent cell-dependent cytotoxicity compared to Erbitux. This indicates that these goats produce an improved cetuximab version with the potential for enhanced effectiveness and better safety profile compared to treatments with Erbitux. In addition, our study validates transgenic goats as an excellent platform for large-scale production of therapeutic mAbs.

Cloning and sequence determination of bovine insulin-like growth factor binding protein-2 (IGFBP-2): Comparison of its structral and functional properties with IGFBP-1

1992 ◽  
Vol 48 (2) ◽  
pp. 215-226 ◽  
Author(s):  
Walker H. Busby ◽  
Robert H. McCusker ◽  
David R. Clemmons ◽  
Maureen J. Bourner ◽  
Ned R. Siegel ◽  
...  
Keyword(s):  
Growth Factor ◽  
Functional Properties ◽  
Binding Protein ◽  
Bovine Insulin ◽  
Insulin Like Growth Factor ◽  
Sequence Determination ◽  
Factor Binding

Cloning and characterization of a cDNA encoding the β-subunit of the bovine insulin-like growth factor-1 receptor

DNA Sequence ◽  
1991 ◽  
Vol 1 (6) ◽  
pp. 405-406 ◽  
Author(s):  
M. Sneyers ◽  
R. Kettmann ◽  
S. Massart ◽  
R. Renaville ◽  
A. Burny ◽  
...  
Keyword(s):  
Growth Factor ◽  
Bovine Insulin ◽  
Insulin Like Growth Factor ◽  
Β Subunit

Large–Scale Production of Proteins from Mammalian Cells

1988 ◽  
Vol 6 (5) ◽  
pp. 518-523 ◽  
Author(s):  
Malcolm Rhodes ◽  
John Birch
Keyword(s):  
Mammalian Cells ◽  
Large Scale ◽  
Scale Production ◽  
Large Scale Production

scholarly journals Insect Cecropins, Antimicrobial Peptides with Potential Therapeutic Applications

2019 ◽  
Vol 20 (23) ◽  
pp. 5862 ◽  
Author(s):  
Daniel Brady ◽  
Alessandro Grapputo ◽  
Ottavia Romoli ◽  
Federica Sandrelli
Keyword(s):  
Infectious Diseases ◽  
Antimicrobial Peptides ◽  
Mammalian Cells ◽  
Large Scale ◽  
Scale Production ◽  
Gram Negative ◽  
Social Burden ◽  
Large Scale Production ◽  
Conventional Antibiotics ◽  
The Cost

The alarming escalation of infectious diseases resistant to conventional antibiotics requires urgent global actions, including the development of new therapeutics. Antimicrobial peptides (AMPs) represent potential alternatives in the treatment of multi-drug resistant (MDR) infections. Here, we focus on Cecropins (Cecs), a group of naturally occurring AMPs in insects, and on synthetic Cec-analogs. We describe their action mechanisms and antimicrobial activity against MDR bacteria and other pathogens. We report several data suggesting that Cec and Cec-analog peptides are promising antibacterial therapeutic candidates, including their low toxicity against mammalian cells, and anti-inflammatory activity. We highlight limitations linked to the use of peptides as therapeutics and discuss methods overcoming these constraints, particularly regarding the introduction of nanotechnologies. New formulations based on natural Cecs would allow the development of drugs active against Gram-negative bacteria, and those based on Cec-analogs would give rise to therapeutics effective against both Gram-positive and Gram-negative pathogens. Cecs and Cec-analogs might be also employed to coat biomaterials for medical devices as an approach to prevent biomaterial-associated infections. The cost of large-scale production is discussed in comparison with the economic and social burden resulting from the progressive diffusion of MDR infectious diseases.

INSULIN-LIKE GROWTH FACTOR I/SOMATOMEDIN C (IGF-I/Sm C): COMPARISON OF NATURAL, SOLID PHASE SYNTHETIC AND RECOMBINANT DNA ANALOG PEPTIDES IN TWO RADIOLIGAND ASSAYS

Endocrinology ◽  
1984 ◽  
Vol 115 (6) ◽  
pp. 2490-2492 ◽  
Author(s):  
DON SCHALCH ◽  
DORIS REISMANN ◽  
CAROL EMLER ◽  
RENE HUMBEL ◽  
CHOH HAO LI ◽  
...  
Keyword(s):  
Growth Factor ◽  
Recombinant Dna ◽  
Solid Phase ◽  
Insulin Like Growth Factor ◽  
Somatomedin C ◽  
Factor I ◽  
Igf I ◽  
Growth Factor I

Characterization and cloning of a bovine insulin-like growth factor-binding protein

1990 ◽  
Vol 5 (1) ◽  
pp. 77-84 ◽  
Author(s):  
F. Z. Upton ◽  
L. Szabo ◽  
J. C. Wallace ◽  
F. J. Ballard
Keyword(s):  
Amino Acids ◽  
Growth Factor ◽  
Binding Protein ◽  
Sequence Similarity ◽  
Bovine Insulin ◽  
Insulin Like Growth Factor ◽  
Protein Precursor ◽  
Factor Binding ◽  
Bovine Cell ◽  
Glycosylation Sites

ABSTRACT The primary structure of the insulin-like growth factor-binding protein (IGFBP) produced by the bovine kidney cell line, MDBK, has been deduced from the cDNA clone. The MDBK binding protein precursor consists of a hydrophobic pre-peptide of at least 26 amino acids and a mature protein of 284 amino acids. The predicted protein sequence shares extensive sequence similarity with both the rat (82%) and human (89%) IGFBP-2s, so that the MDBK binding protein is clearly the bovine counterpart of IGFBP-2. The protein has limited similarity with classes 1 (31%) and 3 (31%) human IGFBPs, except that all 18 cysteine residues are conserved. Other features deduced from the bovine IGFBP-2 cDNA include: an abundance of leucine in the pre-peptide, an Arg-Gly-Asp sequence, absence of N-linked glycosylation sites, and an imperfect polyadenylation signal as well as an ATTTA motif in the 3′ non-coding DNA. Western blotting indicated that this binding protein is widely distributed in bovine fluids as well as in media conditioned by bovine cell lines. Proteins immunologically related to bovine IGFBP-2 were detected not only in sheep, but also in chickens, indicating that this IGFBP is not exclusively mammalian.

scholarly journals Bovine Insulin-Like Growth Factor Binding Protein-3: Organization of the Chromosomal Gene and Functional Analysis of Its Promoter

Endocrinology ◽  
1997 ◽  
Vol 138 (7) ◽  
pp. 2856-2862 ◽  
Author(s):  
N. E. Erondu ◽  
B. Toland ◽  
M. Boes ◽  
B. Dake ◽  
D. R. Moser ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Growth Factor ◽  
Binding Protein ◽  
Bovine Insulin ◽  
Insulin Like Growth Factor ◽  
Chromosomal Gene ◽  
Factor Binding

Production of bovine insulin-like growth factor 2 (bIGF2) in Escherichia coli

Gene ◽  
1991 ◽  
Vol 101 (2) ◽  
pp. 291-295 ◽  
Author(s):  
Alan M. Easton ◽  
James K. Gierse ◽  
Ramnath Seetharam ◽  
Barbara K. Klein ◽  
Claire E. Kotts
Keyword(s):  
Escherichia Coli ◽  
Growth Factor ◽  
Bovine Insulin ◽  
Insulin Like Growth Factor
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