Large Scale Antibody Production and Purification

Successful large scale antibody production and purification 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 large-scale antibody 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 these unfrozen, subcloned cells directly into production. This allows a decrease in the timeline of getting purified antibody into the customer’s hands. Green Mountain Antibodies also accepts external cell lines. We efficiently process hundreds of cell lines per year 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. GMAb can arrange to test any uncertified hybridoma lines prior to production. Maintaining a clean lab is important to us and all of our customers.

When did you last subclone the line?
GMAb suggests routine subcloning of cell lines to select for healthy, antibody producing cells.

What media has been used for cell growth?
Optimal cell growth will best continue using the media formulation to which the cells are accustomed and likely frozen down in. Cells will be transitioned to serum free media to generate antibody production supernatants.

Do I have ample back up of frozen vials?
Be sure to let us know if this is the last vial of this hybridoma line so we can plan to modify media and handling as necessary. We can also plan to freeze down an appropriately sized cell bank.

Cell Culture and Large-Scale Antibody Production and Purification Services:

  • Subcloning and screening of existing cell lines.
  • Cell banking.
  • In vitro production of 50mgs to grams of purified antibody.

In vitro Productions

  • A pilot run will determine the cell line’s production rate prior to scale-up. GMAb produces and purifies a smaller volume of antibody-containing spent media.
  • We can make a production volume recommendation when an estimated production rate (mg/antibody/liter/day) is available.
  • 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, DLS, BLI affinity ranking by Octet/Gator, 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.

Advantages of Large-Scale Antibody Production and Purification at GMAb

  • 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.
  • We can formulate purified antibody in appropriate buffer or lyophilize 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.

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