See also Carbon Filters

Companies:  Merk Group:  Merck KGaA   (subsidiaries: EMD Millipore Corporation)

Separation of Antibodies using Hydrophobic Chromatography Resins

Polystyrene or poly(ethyl)styrene or Vinylbenzene:

–cross-linked with Copolymers od divinylbenzene and ethyleneglycol methacrylate

Skudas (US 15/123110, published as US 2017/0073394) dicloses HCPs, antibody fragments and low MW substances can be separated from antibodies using a hydrophobic chromatography material made of cross-linked vinylbennzene, ethylstyrrene, poly(ethyl)styrene-divinylbenzene or poly(ethyl)styrene-divinylbenzene ethyleneglycol-dimethylacrylate). Preferably the resin is composed of polystyrene or poly(ethyl)styrene, which is cross-linked with copolymer of divinylbenzene and ethyleneglycol methacrylate in a ratio of 98:2 up to 10:90% by weight. The contacting is operated in flow through mode where the HCPs and fragments are absorbed to the hydrophobic chromatography material. 

Separation of Antibodies using Activated Carbon 

 Different Modes used

–Flow through mode: 

Bian (US 13/565463, published as US2013/0197200 and US 9096648; see also US 14/747029, published as US 2016/0016992; see also US 16/358,845, published as US 2019/0218250) discloses that carbonaceous material such as activated carbon can be incorporated into a chromatography column based antibody/ protein purification process in a flow through mode, resulting in reducing the burden of chromatography columns. The activated carbon can be used either upstream or downstream of a capture chromatography step to reduce the level of one or mroe impurities. In one embodiment, an antibody sample is contacted with an affinity media and the flow through contacted with a carbonaceous material such as activated carbon and the eluate from the activated carbon contacted with an anion exchange media. 

Ishihara (US 13/826195, published as US 2014/0046038) dicloses using activated carbon for the separation of proteins such as antibodies in a non-adsorption mode. Examples of the activated carbon include mineral based such as coal based and petroleum based activated carbon. Examples of plant based inlcude wood based activated carbon. The raw material of the activated carbon must be carbonacoues which includes materials such as sawdust, charcoal, ash, peat moss, peat or wood chip, coals such as lignite, brown coal or antracite, coal pitch, petroleum pitch, oil carbon, rayon, acrylonitril or phenol resin. 

Kozlov (US 14/891,724, published as US 2106/0090399) discloses a method of reducing antibody fragments from an antibody prepration by applying the same to activated carbon where the activated carbon binds the fragments and then removing the AC as by filtration or centrifugation. 

—-Protein A -AC (flow through) -AEX (flow through) -CEX

Kozlov (US 2018/0215786) also disclsoes a flow-through process for purifying a target molecule sucha s an antibody from a Protein A eluate which includes the steps fo contacting the eluate form the (a) Protein A column with activated carbon, (b) contacting a flow through from the AC with AEX and (c) contacting the flow through with CEX at a density of about 1-30 mM, wherein the eluate flow continously through steps (a)-(c). 

Singh (US 2013/0012689) discloses a continous profess for purifying a target molecule which includes flow through activated carbon followed by a flow through AEX and CEX media. 

Xenopoulos (US2015/0133636) discloses purification of an antibody/protein by from a Protein A eluate by contacting the eluate with activated carbon and then contacting the flow through with AEX, the flow through from this with CEX and obtaining the flow through sample comrpsing the antibody.

Xenopoulos (US 15/654876, published as US 2017/0320909) also discloses a method of purifying an antibody by precipitating contaminant(s) from a sample and then subjecting the clarified sample to a bind and elute chromatography step that incldues at least two separation units and then subjecting the eluate to a flow through purification step that includes two or more media, one of which is activated carbon and the other which are AEX or CEX. The process is advantageously practiced concurrently for at least a portion of two steps using surge tanks and/or static mixers. In a specific emobdiment the flow-through purificaiotn step is carried out as activated carbon followed by AEX and then CEX with an in-line static mixer and/or surge tank being used between AEX and CEX to change pH.