Protein purification

Equipment

AKTA PURE

Äkta Pure represents the most recent and upgraded version of the Äkta Purifier:

  • ‌New software Unicorn 7 - simplifies the creation of automated programs and calculates protein concentration directly from the chromatogram
  • Equipped with sample pump and air sensor - no need to watch for sample loading, it automatically stops when the sample is over
  • Capable of multistep programs - save time and improve protein purity by coupling affinity chromatography and gel filtration in a single run

Äkta Pure is compatible with any type of column and with purification programs requiring a maximum of 4 different buffers. The instrument works at room temperature.

The use of Äkta Pure requires no training, as the instrument is operated by the technician. Users are responsible for sample preparation, but attending to the purification is not necessary.

 

 

AKTA START

Äkta Start is used for purification protocols requiring a maximum of two different buffers. Automated programs are available for the fast purification of His-tagged proteins and antibodies, and to perform buffer exchange of protein samples. Customised programs can also be developed upon request. 

The instrument is controlled by a user-friendly software (Unicorn Start). This allows researchers to work autonomously, after receiving an induction.

Äkta Start can work both at room temperature and at 4°C. This instrument is compatible with chromatography columns giving a max. back-pressure of 0.5 MPa. 

 

 

COLUMNS

Researchers are free to bring their own column or borrow one of the facility columns at no extra cost. A list of facility columns currently available is provided.

‌If you wish to use your own column, please remember that:

  • HPLC columns are not compatible with Äkta systems
  • Äkta Start is compatible with columns giving a max. back-pressure of 0.5 MPa (pressure values are typically reported on the datasheet of the column)

If you are not sure whether your column is compatible with one of our systems, please contact the technician for advice.


New users

All users are encouraged to use the Äkta Start whenever possible. Permission to book the instrument will be granted after receiving the necessary training. A training session takes approximately 1 hour, and covers the basics to use the instrument: prepare the sample, prime the machine, start a purification program, export your data.

Please contact the technician to book a training session.

No training is required to use the Äkta Pure, as the instrument is always operated by the technician. Researchers will be given booking permission upon request, after being informed about the requirements for sample and buffers preparation.


Booking guidelines

Reservations of Äkta Start do not require approval. The instrument can be booked out-of-hours, but assistance is provided only during working hours. If you need assistance throughout the purification, please inform the technician as soon as possible.

Reservations of Äkta Pure are allowed only during working hours and are subjected to approval, as you always need assistance to operate the machine. Out-of-hours bookings will be accepted only on exceptional circumstances.

If you wish to use a Gel Filtration column: you can do that only on Äkta Pure. Gel filtration requires several hours, so please book a full working day (from 9.00 am to 5.00 pm).

If you wish to purify at 4°C: you can do that only on Äkta Start. Please contact the technician to find a suitable time slot for the reservation, as the machine will not be available to other users for the entire day.

Reservations are managed through the Institute’s Booking System. A minimum booking notice of 12 hours is required for the Äkta Start, and 18 hours for the Äkta Pure. 

When you make a reservation please book a minimum of 2 hours, as time is required also to prepare the instrument for the run, and to clean afterwards.

If you wish to borrow one of the facility columns, please write the name of the column on the reservation.


Charges

The facility runs as a not-for-profit service. Charges cover the cost of consumables and maintenance of instruments:

  •         Äkta Start: £12/hour
  •         Äkta Pure: £25/hour

For gel filtration only, the hourly rate is replaced by a fixed charge of £70. The use of the column requires the booking of the Akta Pure for a full working day (9.00 am-5.00 pm), during which a maximum of 2 runs are allowed.   

Cancellation policy: no charge for cancellations (any time before the reservation starts). Full charge applies if the user doesn't cancel and doesn't show up. Charges may be waived under exceptional circumstances.        


Publication policy

As an academic core facility, the Protein Production Service in common with other universities follows the internationally established guidelines stipulated by journals and publication houses. Please acknowledge any facility contribution to your research by following the guidelines below:

  1. All publications resulting from the use of instruments within the facility should acknowledge the Protein Production Service as a whole, e.g. ‘the authors gratefully acknowledge the Protein Production Service for their support & assistance in this work’
  2. If a staff member of the Protein Production Service provided significant help or generated additional data personally, the staff member should be acknowledged by name, alongside the facility if applicable, e.g. ‘the authors thank *** of the Protein Production Service for their support & assistance in this work’
  3. If a staff member of the Protein Production Service contributed more than just routine techniques (e.g. development or adaptation of protocols to suit samples or materials, (re)designing experiments, extensive data analysis and interpretation), it constitutes contributions meriting co-authorship in papers.

The Protein Production Service staff expect to be informed of any potential publications resulting from their work. The staff should have the opportunity to participate in drafting the pertinent part of the paper and give final approval to the wording and conclusions drawn before publication. If you are uncertain about co-authorship or have any questions or concerns about this, please discuss this issue with the facility manager in advance.

Please send a reprint of the paper, or an e-mail including the reference information for the publication, to the Protein Production Service. We also politely request to acknowledge the Protein Production Service in your presentations and to cite the facility in grants, as appropriate.