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Purification of recombinant human growth hormone (rhGH) on multimodal chromatography media

IP.com Disclosure Number: IPCOM000219441D
Publication Date: 2012-Jun-29
Document File: 5 page(s) / 134K

Publishing Venue

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Abstract

Recombinant human growth hormone has been successfully purified on the multimodal ion exchange resins CaptoTM adhere ImpRes and CaptoTM MMC ImpRes using ethanol/salt gradients (Capto adhere), decreasing pH gradients (Capto adhere) and increasing pH gradients (Capto MMC).

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Purification of recombinant human growth hormone (rhGH) on multimodal chromatography media

Human growth hormone from fermentation broths can be chromatographically purified on multimodal ion exchange media such as e.g. CaptoTM adhere ImpRes and Capto MMC ImpRes (available from GE Healthcare) with high selectivities using elution with ethanol/salt or pH gradients.

Example 1 - purification on Capto adhere ImpRes with ethanol/salt elution

6 micrograms of rhGH was applied to a Tricorn 5/50 column packed with Capto adhere ImpRes, which had been equilibrated with a 50 mM Tris buffer pH 7.4 containing 10% ethanol (A buffer). The column was eluted with a B buffer consisting of 50 mM Tris buffer pH 7.4 containing 50% ethanol and 500 mM NaCl. The elution gradient was linear from 100% A buffer to 100% B buffer over 10 column volumes.

UV 1_280_Chrom.1:Capto Adhere ImpRes 10ul grad... UV 2_220_Chrom.1:Capto Adhere ImpRes 10ul grad... Cond_Chrom.1:Capto Adhere ImpRes 10ul grad 10c... Conc B_Chrom.1:Capto Adhere ImpRes 10ul grad 1... pH_Chrom.1:Capto Adhere ImpRes 10ul grad 10cv ... Injection_Chrom.1:Capto Adhere ImpRes 10ul gra...

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Figure 1. Chromatogram of rhGH on Capto adhere ImpRes, eluted with ethanol/salt.


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Example 2 - purification on Capto adhere ImpRes with decreasing pH gradient

6 micrograms of rhGH was applied to a Tricorn 5/50 column packed with Capto adhere ImpRes, which had been equilibrated with a 50 mM Tris buffer pH 7.4 containing 10% ethanol (A buffer). The column was eluted with a B buffer consisting of 50 mM sodium acetate buffer pH 4.0 containing 10% ethanol. The elution gradient was linear from 100% A buffer to 100% B buffer over 10 column volumes.

UV 1_280_Chrom.1:Capto Adhere ImpRes 10ul hGH p... UV 2_220_Chrom.1:Capto Adhere ImpRes 10ul hGH p... Cond_Chrom.1:Capto Adhere ImpRes 10ul hGH pH gr... Conc B_Chrom.1:Capto Adhere ImpRes 10ul hGH pH ... pH_Chrom.1:Capto Adhere ImpRes 10ul hGH pH grad... Injection_Chrom.1:Capto Adhere ImpRes 10ul hGH ...

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Figure 2. Chromatogram of rhGH on Capto adhere ImpRes, eluted with decreasing pH gradient.


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Example 3 - purification on Capto adhere ImpRes with decreasing pH gradient, no ethanol

6 micrograms of rhGH was applied to a Tricorn 5/50 column packed with Capto adhere ImpRes, which had been equilibrated with a 50 mM Tris buffer pH 7.4 (A buffer). The column was eluted with a B buffer consisting of 50 mM sodium acetate buffer pH 4.0. The elution gradient was linear from 100% A buffer to 100% B buffer over 10 column volumes.

UV 1_280_Chrom...

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