Miscellaneous Antibody Services
Characterization | Conjugation | Purification | Banking and Preservation | Mycoplasma Testing and Decontamination
Antibody Characterization Services
To ensure that a purified antibody will work to its best potential, Maine Biotechnology has assembled a group of diagnostic tools that when used together provides antibody characterization data. This data provides not only basis for selection of appropriate storage parameters but also a means to track the consistency of each production.
Size Exclusion Chromatography (SEC): Allows us to determine percent purity of an antibody as well as to calculate and differentiate the percentage of monomeric, dimerized, aggregated and fragmented antibody.
IsoElectric Focusing (IEF): Determines the isoelectric point of an antibody, loosely translated to the pH of the antibody. It also reveals isoforms or variants in the purified material.
Enzyme Linked Immunosorbant Assay (ELISA): If the antigen is provided, the staff at MBS can also run a standard ELISA on the purifed antibody to validate that it binds to the antigen.
SDS-PAGe, reduced and Non-reduced: The SDS-PAGe is supportive of the SEC and can be used to validate observations made in the non-reduced antibody. Upon reduction the antibody is dissociated into heavy and light chains, and this further characterizes the structural integrity of the purified antibody.
Conjugation
Conjugations include chromatographic removal of unconjugated ligand. Approximate conjugate strength is assayed by ELISA. A final Quality Assurance report is sent with the conjugated material. Custom Conjugations include:
· Murine Monoclonal Antibody and Antigen Conjugation
· Conjugation to HRP
· Conjugation to Biotin
· Conjugation to FITC
· Conjugation to Phycoerytherin
Purification
Ascites or In Vitro Purification
Murine monoclonal IgGs are purifed from ascites or in vitro technology by protein A or G affinity chromatography. MBS purifications are eluted by pH gradient to ensure the gentlest conditions possible. (read more about our purification methods in News section)
· Purification to yield final antibody amounts
· Concentration/dialysis to desired transport buffer
· Testing of puried antibody for protein content, Ig concentration
· Sterile filtration of final product
· Addition of NaN3 upon request
· Percent Purity determined by Size Exclusion Chromatography allowing
differentiation between monomeric, dimerizedaggregated, and fragmented antibody.
· IsoElectric point determination
Click here to initiate your purification with the Purification Service Request Form.
Polyclonal Purification
Rabbit, goat or sheep antiserum are purifed by Protein A of G affinity Chromatography. Most serum yields between 5 to 10 mg/mL Ig.o Affinity purification also available.
Affinity purification Services
Antibodies and Antigens can be purified by affinity chromatography. MBS will prepare a column using your antibody/antigen coupled to Sepharose 4B. Your antibody/antigen will then be purifed after elution from this specially prepared chromatography column. An Organic Chemist will work directly with you to identify the appropriate chemistry for your column.
Cell Banking and Preservation Services
Be sure your cells are safely banked and protected. MBS will expand cell lines for desired vial storage numbers. MBS will create and maintain a Master Cell Bank for your cells and perform viability testing as needed per cell line.
Mycoplasma Testing
Mycoplasma testing service is done by DNA Fluorochrome and Ultraviolet microscopy to reveal any extranuclear staining characteristic of mycoplasma contamination.
Results from testing are given as positive, negative, or questionable.
1 vial of frozen 1 x 10^6 cells are needed.
Mycoplasma Decontamination
Cells can be "cleaned-up" by retrieving cells after an In
Vivo passage.
Cells are injected into IFA primed mice
Retrieval of cells from mice after 10-20 days growth In Vivo
Cells are rescreened for mycoplasma
Cells are reestablished In Vitro
Cells are subcloned to reestablish monoclonality