There is no kit to make ADC.
There are however simple techniques that you can use. Key thing however is the chemistry that you use to do that. Let's assume maleimide chemistry, which is the most common one nowadays.
You have to have a toxin with appropriate linker (such as MMAE and maleimide) and appropriate conditions to conjugate it to an antibody. Then you have to purify, as you will have a lot of various species, so called DARs (Drug Antibody Ratio). DAR 0 is an antibody with no payload, DAR 1 is an ADC with one toxin and so on...
Depends what you want to do really. Do you want to to see efficiency of your reactions, do you want to see if your antibody can be conjugated, do you want to produced some ADCs and use them for something?
One of ways to detect ADCs is analyse them by HIC (hydrophobic interaction chromatography), but that depends on the toxin and its hydrophobicity.
We really want to screen our antibodies in vitro to determine whether they could be used for ADC. We don't care about the DAR as long as the DAR is higher than 1.
If I understand correctly you have few antibodies, you want to make ADC from them and quickly screen it. Or is it the other way round. That you want to screen your antibodies to then be able to know if they are ok for ADC.
In first example, you won't find a kit, because it is not such a simple task.
Conjugation is a key, which depends on many factors.
In the second case I think it is more of decision whether an antibody is a good ADC candidate or not. This decision is based on what receptor this antibody binds to, is this receptor present on cancer cells or also in normal cells. In vitro studies should focus on measuring binding of IgG to the cell, e.g. by having bacteria with overexpressed receptor to which this antibody binds and of course a control with different receptor. You can scree antibodies and see which one binds the best.
I would say that the key thing is specificity of antibody towards receptor.
We have dozens of antibodies which can bind to the surface of several cancer cell lines. However, we don't know the targets of these antibodies. So we would like to make ADCs to screen them.
Conjugation with fluorescent dye (FITC or Alexa), adding to positive cell lines and then imaging using confocal microscopy will indicate if ADC internalises and where. The key step is conjugation and the synthetic chemistry of linker-drug. However, you would have to purify them to some extent, e.g. to remove non-reacted drug. Although the principle is simple it is not as straightforward, that is why there are so far no kits available.
Hi Huiwu Zhao and Kosma Jurlewicz just wanted to say I work with a UPenn-based biotech company AlphaThera and we currently provide ADC conjugation reagents for MMAE drug. These reagents support conjugation with a wide variety of antibodies and buffers. If you are interested check out the product here: https://www.alphathera.com/antibody-drug-conjugation-kits/mmae
The conjugation is site-specific and directly to the heavy chain of antibodies. Some benefits include 30-second hands on time for conjugation, precise control over your cell-killing assays, and widely compatible. We will be adding more drugs to the product line as well.
lysine 5' method can be used to form antibody-drug bioconjugate, This video has a link of a commercial kit in its description which can help you I hope. https://www.youtube.com/watch?v=SJEulFA2m5c&list=PLTCBtzqkKa4QhazLdinhoIuIx-HV6YDCX … Read more
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