If you are following a standard protocol, you might expect to collect around 3 to 5 ml of eluted sample, but this can vary. It's best to consult the specific protocol you are following or the manufacturer's instructions for the column.

Hi,

The final sample volume that can be effectively processed through these columns typically ranges from 0.5 mL to 1 mL. You can use purification on various antibody sources, such as serum and cell culture supernatants etc.

Regards,

Amal.

There is a simple and a complex answer:

Simple: In most cases, 2 mL are suitable.

Complex: You have to know your priorities: maximum yield, maximum concentration or maximum activity of the antibodies.

In the first case, you have to collect up to 5 mL or even more to get as much antibody as possible, however, then the solution is relatively diluted.

In the second case, you try to collect only the highest part of the elution peak and discard or recycle the rest. Your yield may be only 50%, but the concentration is high. You also can use a harsher elution condition (lower pH) as usual to obtain a sharper elution peak. This however may reduce the activity of your antibody.

In the third case, you elute your columns with a higher pH (has to be tested) as usual to maintain the maxium activity of the antibodies. But then the elution curve may be broader and the solution is even more diluted.

My suggestion: Take a microwell plate (ideally UV transparent) and collect the eluate in fractions of about 3 droplets. After measuring the absorbance in UV (280 nm), you can decide, which fraction you take and which fractions you may put together to get your final antibody solution. However, do not keep too many fractions.