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Recombinant Protein Expression in Mammalian Cells

Application list of expression system Expression system Advantage Fast; Widely used. Fast; Partial post-translational modification Soluble protein; Translation modification similar to mammalian system Better post-translation modification; Inclusion; Difficulty in expression of macromolecular weight proteins High-sugar dew retouching High cost; Long cycles; Recommended Expressions Cytokines; Cytokines; Enzymes Se...

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Bacterial Protein Expression

Introduction Protein Expression in Bacteria Bacterial expression system is an ideal protein expression system of all time because of its nature. It often acts as the first choice in producing recombinant proteins and antibodies. Many bacteria have already been well characterized and investigated until now. Among the huge number of bacteria, E. coli and B. subtilis cell lines are the most commonly used hosts for recombinant protein expression. They replicate extremely fast under the right condition at a low cost, while the expression level is high, which makes it an efficient and economical way for large scale production. Their genomes have been entirely sequenced and the genetic backgrounds and metabolic pathways are well known. Additionally, because of the simple structures, bacteria are more easily transfected than eukaryotes. These desirable characteristics make the expression procedure easier and quicker. But every coin has two sides. Proteins with few modifications could be expressed in bacteria easily. While there is no nucleus, Golgi apparatus, and endoplasmic reticulum in bacteria. These structures are nece...

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Protein Isoform Detection Service

Protein Isoform Detection Service What is Protein Isoform? A protein isoform, or "protein variant", is a member of a set of highly similar proteins that originate from a single gene or gene family and are the result of genetic differences. While many perform the same or similar biological roles, some isoforms have unique functions. A set of protein isoforms may be formed from alternative splicings, variable promoter usage, or other post-transcriptional modifications of a single gene; post-translational modifications are generally not considered. Through RNA splicing mechanisms, mRNA has the ability to select different protein-coding segments (exons) of a gene, or even different parts of exons from RNA to form different mRNA sequences. Each unique sequence produces a specific form of a protein. The discovery of isoforms could explain the discrepancy between the small number of protein-coding regions genes revealed by the human genome project and the large diversity of proteins seen in an organism: different proteins encoded by the same gene could increase the dive...

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Western Blot Analysis

Introduction Many biological and medical diagnostics laboratories employ Western blotting (a.k.a. immunoblotting) to detect proteins from cell extracts, blood serum or saliva sputum samples. Western blotting is considered a mature technology, but remains extensively used by researchers in the biotechnology, pharmaceutical and diagnostics industries. Customers continue to demand improvements in ease-of-use, speed, and sensitivity, as well as, a better ability to quantify results. Western blot analysis can detect one protein in a mixture of any number of proteins while providing you with information about the molecular weight of the protein. This method, however, is dependent on the use of a high-quality antibody directed against a desired protein. This antibody is used as a probe to detect the protein of interest. Western Blot Analysis Services · Extraction of total proteins from tissues/cells provided by Leading Biology or customers · Constructions of Western blots using total proteins · Immunostaining blots with antibodies provided by customers using ultrasensitive chemiluminescent ECL detection ...

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Protein Expression in Bacteria

Introduction Protein Expression in Bacteria Bacterial expression system is an ideal protein expression system of all time because of its nature. It often acts as the first choice in producing recombinant proteins and antibodies. Many bacteria have already been well characterized and investigated until now. Among the huge number of bacteria, E. coli and B. subtilis cell lines are the most commonly used hosts for recombinant protein expression. They replicate extremely fast under the right condition at a low cost, while the expression level is high, which makes it an efficient and economical way for large scale production. Their genomes have been entirely sequenced and the genetic backgrounds and metabolic pathways are well known. Additionally, because of the simple structures, bacteria are more easily transfected than eukaryotes. These desirable characteristics make the expression procedure easier and quicker. But every coin has two sides. Proteins with few modifications could be expressed in bacteria easily. While there is no nucleus, Golgi apparatus, and endoplasmic reticulum in bacteria. These structures are nece...

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Unique Protein Production Service

Introduction UPP is an expression cell line which could express foreign protein efficiently. The foreign protein can account for more than 20-30% of the total protein it produced, and its principle is based on an enzyme called Marzf that can hydrolyze host mRNA specifically. This enzyme can specificity recognize the ACA sequence of mRNA, and through codon optimization, remove this sequence from the sequence which encoding exogenous protein, and this could avoid the target protein mRNA having the same fate with the host cell protein mRNA. Under the effect of these enzymes, most of the genetic information of the host genes is shielded, while the transcription and translation system still working, this modification can efficiently express the exogenous protein without other interferences. The high Signal-to-noise ratio (SNR) of this technology made the truly high flux protein production came true. With the constant improvement of the system and related technology innovation, by now, we can use this system to achieve the preparation of 90% of proteins, and with the optimization of the production process, we can achieve 40 times concentrated fermentation system for ...

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Cell-Free Protein Expression System

Service Introduction The Cell-free expression system is an external system based on exogenous DNA or mRNA as template, using cell extract which contains cell synthesize organ, protein folding factor and other related enzymes, by adding amino acids and T7 polymerase to achieve the in vitro protein expression system, including the eukaryotic cell-free expression system and the prokaryotic cell-free expression system. Advantages of Cell-Free system 1. Faster data generation: Functional protein can be obtained in only 1 hour, while the cell-based system requires several days. 2. Time-saving: The expression protein can be used directly in the follow-up experiment, no additional purification needed. 3. It is more suitable for the expression of kinases and other toxic proteins, can be marked with modified tRNA, and add unnatural amino acids at specific sites. 4. One system, multiple applications: Proteins expressed can be used in protein-protein interaction experiments, protein-nucleic acid interaction experiments, and protein modification analysis. Leading Biology cell-Free expression system process ...

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Rich in Disulfide Bond Protein Expression

Service introduction As we all know, the structure of the protein which rich in the disulfide bond is relatively complex, the solubility of the target protein expressed by the prokaryotic system is quite low, and is very easy to form an inclusion body, thus, the probability of getting the active protein is very low. Leading Biology using a new prokaryotic expression system, greatly increased the probability of disulfide bond rich protein expression in Escherichia coli, the expression rate is greater than 90%. Service process Service Items Time(weeks) Protein analysis and codon optimization Before project start Gene synthesis 2-4 Vector construction 1 E.coli expression strains transformation and screening 2 Target protein inducible expression ...

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Protein Expression in Mammalian Cells

Application list of expression system ProkaryoticEukaryoteEukaryoteEukaryoteAdvantage Fast; Widely used. Fast; Partial post-translational modification Soluble protein; Translation modification similar to the mammalian system Better post-translation modification; Limitations No post-translation modification; Non-human-derived Glycosylation; Long cycle Lack of partial glycolytitization Recommended Expressions Cytokines; Cytokines; Enzymes Secretion proteins; Kinases ...

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Insect Baculovirus Expression Vector System

Insect baculovirus expression vector system introduction Insect baculovirus expression vector system (BEVS) is a system in which baculovirus worked as a vector of foreign genes, by infect insect cells or insect larvae to produce exogenous proteins. It is an important part of the eukaryotic expression system. Advantages of insect baculovirus expression system 1). Safe 2). Large capacity of carrying foreign genes 3). High expression level of exogenous protein 4). The expressed eukaryotic gene products can be properly folded and modified, thus have good bioactivity. Advantages Compared to mammalian cell expression system: 1). The culture conditions are relatively simple and do not require the CO2 2). The cost is relatively reasonable 3). Not easy contaminated by bacteria 4). High protein content The technical advantages of Leading Biology 1. We use classic Bac-to-Bac technology to create a fast, simple and efficient baculovirus expression platform. With the technique, we can produce recombinant virus quickly, and do not require tedious plaque assay, which greatl...

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