ProteoCool Pills#16: Loading small sample volumes on AKTA instruments

AKTA instruments (as AKTA-FPLC, AKTA-PRIME, AKTA-Purifiers, AKTA-Go, AKTA-pure) represent in the last 30 years the point of reference for protein/antibody purification in both research, development and pharmaceutical production departments since they are robust, easy to use and provided with a software (the Unicorn)  which is a reall uses friendly software.

On the contrary, their application for protein/antibody analytics characterization is mainly limited to some specific columns and approaches (eg SEC chromatography with the Superdex 10/30 or 5/15 coloumns) in those laboratories that do not have budget dedicate to acquire an analitical HPLC. 

 In those case the loading and injection of a small sample volume in reproducible way it a prerequisite.

How we can inject small volumes in an FPLC or other AKTA instruments? 

For volumes between 200-300µl to 1ml,  the standard injection port could be used with an 1ml insulin plastic syringe (with out needle)

For volumes <250µl ,  the standard injection port need to be replaced with a injection filling port designed for small volumes and use an more precise Hamilton syringe (250µl or 100µl volume) for sample injection.  

Until some years ago (as I already wrote to answer to a question in Research gate in 2019) https://www.researchgate.net/post/Loading-small-sample-volumes-on-FPLC), Citiva bioscience (previously GE bioscience) supplied for small sample injection a metal injection filling port (CITIVA cod. 19768701) supplied with a plastic syringe stand suitable for injection of small volumes using Hamilton syringes

Recently those Injection fill port was discontinued and replaced by a plastic filling post (Fill Port, INV-907 cod.18112766)

Also if this filling port, is not supplied with any syringe stand, it could be used directly with an hamilton syringe, as you can see from the following imagine:

Of course to inject so small volumes you need to select also the appropriate:

•          Injection loop (with a small volume)
•         Syringes (with BLUNT tips:

Suggested:

50-100ul sample:  Hamilton 1710N volume 100 μL, needle size 22s ga, needle L 51 mm; Sigma-Aldrich cod. 28634-U cost ~100€;

250-100ul samples: Hamilton® 1725N, volume 250 μL, needle size 22 ga (blunt tip), needle L 51 mm; Sigma-Aldrich cod. 28636-U cost ~80€;