Our recombinant proteins can be used as bio-markers, diagnostic test proteins and also as excellent antigens, producing a strong immune response which yield antibodies in 99% of cases.

Examples                                  Mammalian Targets – Cytokines, Receptors, Enzymes
                                                    Bacterial Targets – TB, Yersinia pestis
                                                    Viral Targets – Hepatitus B, Sars, Vaccinia

Accompanying Services       Polyclonal Antibody Production
                                                    Monoclonal Antibody Production
                                                    Antibody Sequencing
                                                    Antibody Engineering


Please enquire for more details on this service and pricing with our technical services team at  technicalservices@fusionantibodies.com or call us on  +44(0) 28 9043 2800 

Technical Summary
 
Bacterial Recombinant Protein Expression

a) Identification of up to 4 regions within the protein using bioinformatics:
Using in-house bioinformatics packages the target DNA sequence is analyzed, from which up to 4 regions are selected. These regions when synthesized into recombinant protein will produce antigenic fragments. If a specific region is required, antigenic fragments can be designed around that region. The customer can be kept up-to-date with progress at each stage.

 b) Design of PCR primers to amplify these regions:
Primers are designed around the regions identified in stage (a) and ordered. The primers are received and the targeted regions are amplified by polymerase chain reaction (PCR) from the template DNA (supplied or purchased). The PCR is performed using a number of protocols simultaneously including Touchdown, Gradient & at optimal annealing temperatures.

c) Cloning into our in-house expression systems:
The amplified regions are then gel purified ready for the cloning stage.
The PCR products of the amplified regions are restricted, ligated into our vectors and transformed into E. coli. The positive clones are screened for expression performance. 

d) Sequencing and Alignment Analysis:
The positive expressing clones are checked for integrity by DNA sequencing to ensure that the clones contain the correct sequence chosen at stage (a). The results are sent to the customer via e-mail.

e) Identification of relevant expression clones:
Colonies are picked and screened for expression in a high-throughput 96-well plate format. Positive expression clones are identified by SDS-PAGE analysis and western blotting. Growth conditions are further optimised for the clones with the highest expression yields.

f) Scale up of expression:
Further optimisation of growth conditions and additives is performed in a large scale flask to achieve optimal expression yields of recombinant protein.

g) Purification to > 94% purity:
Purification of the recombinant protein by various affinity chromatographic techniques is optimised to give the highest possible yield. A certificate of protein analysis is provided with the sample.

-Stable and transient expression
-Expression of large multimeric proteins
-More complex post translational modifications
-Production functionally active proteins
-Disulphide bridges
-Glycosylation