Successful monoclonal antibody production requires skillful cell culture. At Green Mountain Antibodies, our cell culture expertise is an art and an important part of our technical operations. In vitro production is done via roller bottles at scales of 1 to 20L.
A significant advantage of monoclonal antibodies over polyclonal antibodies is the ability to produce unlimited quantities of the antibody with a single specificity. Monoclonal production also limits lot to lot variability that can be seen with antibodies purified from polyclonal serum. GMAb has designed our production and purification procedures to attain higher antibody yields with reproducibility. Antibodies being used as critical reagents must be held to this standard of quality.
A large percent of GMAb’s in vitro production capacity consists of productions coming directly from our hybridoma development projects. We move subcloned cells directly into production, without ever being frozen. This allows a decrease in the timeline of getting purified antibody into the customer’s hands. Green Mountain Antibodies also accepts external cell lines. Hundreds of cell lines per year are efficiently processed through our labs.
Considerations when sending external cell lines
Are the cells free of mycoplasma?
When sending Green Mountain Antibodies a cell line, please enclose documentation certifying that the vial of frozen cells is mycoplasma free. If the hybridoma line has not been tested, GMAb will arrange to test the cells prior to starting the production. Maintaining a clean lab is important to us and all of our customers. Mycoplasma Contamination Policy
When was the line last subcloned?
GMAb suggests routine subcloning of cell lines to select for healthy, antibody producing cells.
What media has been used for cell growth?
Using the media formulation the cells are accustomed to, and likely frozen down in, is best to continue with optimal cell growth. Cells will be transitioned to serum free media to generate antibody production supernatants.
Do I have ample back up of frozen viles?
If this is the last vial of this hybridoma line, please share with GMAb so extra care can be taken. We can also plan to freeze down an appropriately sized cell bank.
Cell Culture and Antibody Production Services:
- Subcloning and screening of existing cell lines
- Cell banking
- In vitro production of 50mgs to grams of purified antibody
In vitro Productions
- The cell line’s production rate is determined prior to scale-up, via a pilot run. GMAb produces and purifies a smaller volume of antibody-containing spent media.
- Once an estimated production rate (mg antibody/liter/day) is available, a recommendation of production volume can be made.
- The GMAb approach leads to earlier intervention if there is significant decrease or increase in antibody production levels.
- The use of serum-free media decreases possible contamination of purified antibody by bovine immunoglobulin.
- If the final antibody needs to be endotoxin low, our process can accommodate this.
- Subcloning the cell line before large scale production results in improvement of antibody yields and lower cost.
- Purified antibody is provided in sterile phosphate-buffered saline. Other solvent options and characterization are available; such as SDS-PAGE, DLA, Octet affinity ranking, ELISA, and DNA sequencing.
- When the hybridoma cell line is in culture for production, this is an opportunity to further preserve the line with cell banking services and by sequencing the antibody variable regions.
- Obtain mg to gram amounts of purified antibody
- Production in culture eliminates mouse ascites serum immunoglobulins
- Production can begin immediately after subcloning, saving process time and a freeze cycle
- Serum-free conditions lowers contamination of bovine immunoglobulin
- Purified antibody can be formulated in appropriate buffer or lyophilized so you can use immediately
- Rigorous characterization of the purified antibody helps to protect against lot to lot variability
Request a Quote
To initiate a production, please contact us for a quote. GMAb requests certification that the hybridoma cells are free of mycoplasma, an estimate of production rate, and any information you have on growth conditions including doubling times, media requirements, and antibody isotype.