The Purification Insights Blog

QuickTip #6: Use true buffers

Posted by The Bio-Works Team on Dec 4, 2019 9:15:00 AM

A critical and fundamental factor in both analytical and preparative chromatography is the use of true buffers. In other words, mobile phases with buffering capacity. For example, in peptide purification, there are concentrated bands of various components of the sample, including the target peptide, moving through the column and you need to have control over their tendency to dissociate...

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Topics: Quick Tips, Tips and Tricks, Protein purification, Peptides, Buffers

QuickTip #5: Filter everything

Posted by The Bio-Works Team on Nov 26, 2019 11:45:00 AM

Filtering is crucial in order to protect the packed column from all solid particles in feeds or eluents. Clogging of the column will increase back-pressure and disturb the feed loading zone causing reduced resolution and it is a common problem...

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Topics: Quick Tips, Tips and Tricks, Ion exchange chromatography, IEX, Resin fouling, IMAC, Filter, SEC

Ten things you must know when producing synthetic therapeutic peptides

Posted by The Bio-Works Team on Nov 18, 2019 9:40:39 AM

Peptide therapeutic agents are enjoying increasing success but they are challenging to produce and, not least, to purify. Purity requirements are, in fact, approaching those for small molecules. Synthesis generates a challenging crude mixture containing failed sequences and chemical variants. 

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Topics: Tips and Tricks, Biologics, Biopharmaceuticals, Peptides, Peptide drugs, orthogonal, synthetic peptides

QuickTip #4: yield of His-tagged proteins

Posted by The Bio-Works Team on Nov 14, 2019 9:15:00 AM

Immobilized Metal Ion Affinity Chromatography (IMAC) is used for purification of His-tagged proteins. Ni-NTA based resins are the most commonly used as the Ni2+ ion is considered to have the strongest affinity to histidine-tagged proteins but stringent requirements on the yield of target protein can make it well worth the extra effort of trying another metal ion to optimize the purification process...

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Topics: Quick Tips, Tips and Tricks, Protein purification, His-tagged, IMAC

QuickTip #3: optimizing mAb purification

Posted by The Bio-Works Team on Oct 14, 2019 1:38:24 PM

Designing an optimized purification process means maximizing yield and purity, and at the same time controlling costs. So, you remove a lot of bulk impurities during the first steps, and focus on high yield steps at the end, when every gram is precious. If you’re purifying mAbs, it would also be good to remove impurities before affinity chromatography to protect the protein A column and reduce the need for cleaning in place (CIP).

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Topics: Quick Tips, mAbs, Tips and Tricks, Ion exchange chromatography, guard column, Multimodal

QuickTip #2: choosing IEX resin

Posted by The Bio-Works Team on Sep 12, 2019 11:55:00 AM

Recovery of the biological activity of your target substance is a priority, and it’s important to check for its pH and ionic strength stability values...

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Topics: Quick Tips, Tips and Tricks, Ion exchange chromatography, IEX

QuickTip #1: cleaning IEX columns

Posted by The Bio-Works Team on Sep 5, 2019 3:23:49 PM

When an IEX resin has been used for some time, impurities may build up in the column causing loss of resolution or increase in back pressure (shown on the pressure monitor or seen by gap formation on the top of the column)...

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Topics: Quick Tips, Tips and Tricks, Ion exchange chromatography, IEX, Cleaning in place

Some general tips on protein purification strategy

Posted by The Bio-Works Team on Apr 8, 2019 4:06:46 PM

Can you generalize about something as broad as protein purification strategy? Surely it depends on the protein and the situation in general? Well, yes, of course. But there are some general aspects which might be valuable to think through before setting up any specific protein purification.

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Topics: Tips and Tricks, Protein purification

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