Precipitation of biomolecules with negatively charged polymers

Details Precipitation of biomolecules with negatively charged polymers Precipitation of biomolecules with negatively charged polymers

C - Chemistry, Metallurgy

07

K

C07K 1/32 (2006.01) B01D 15/08 (2006.01) B01D 21/01 (2006.01) C07K 16/00 (2006.01)

Patent

CA 2748809

The present invention relates to methods of isolating biomolecules. More particularly, the invention relates to methods for isolating antibodies (mAbs) and related proteinsincluding antibody fragments (Fabs) under conditions where they are positive and relatively hydrophobic and will react with negatively charged polymer to form polymer-protein complexes which precipitate. The isolation can be accomplished usinginexpensive and biocompatible negatively charged polymers such as polyacrylic acid or carboxymethyldextran polymers of various molecular weights as precipitant.Itoccurs at relatively high concentrations of polymer (e.g. 10%) and high salt concentration ( > 50mM) and conductivity (e.g. > 10 mS/cm) over wide range of pH.

La présente invention concerne des procédés permettant d'isoler des biomolécules. L'invention concerne plus particulièrement des procédés permettant d'isoler des anticorps (mAbs) et des protéines apparentées, notamment des fragments d'anticorps (Fabs), dans des conditions où ceux-ci se trouvent à l'état positif et relativement hydrophobe et sont donc aptes à réagir avec des polymères chargés négativement pour former des complexes polymère-protéines qui précipitent. L'isolement peut s'effectuer en utilisant, comme agents de précipitation, des polymères chargés négativement, biocompatibles et bon marché tels que de l'acide polyacrylique ou des polymères du carboxyméthyldextrane de différents poids moléculaires. La précipitation survient à des concentrations relativement élevées de polymère (par exemple 10 %), avec une concentration saline élevée (> 50 mM) et une conductivité élevée (par exemple > 10 mS/cm), sur une plage étendue de pH.

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