GeneMedi's products and data of Antibody-drug conjugate (ADC)

Product list

Case study: Product data of Antibody-drug conjugate (ADC)

Antibody	Payload	QC
Ab-001	VCMMAE	SDS-PAGE(reducing and non-reducing)
Human IgG1 Control		DAR
Ab-002		Cytotoxity assay

First, we used a higher dose of reducing agent (TCEP) and small toxic molecule (vcmmae) to ensure the success of coupling. SDS-PAGE (reducing DTT & non reducing DTT) results showed that we successfully linked MMAE to the antibody. Here you are the conjugate manufacturing.

1. human IgG1-ADC: SDS-PAGE (reducing) and SDS-PAGE (non-reducing)

SDS-PAGE(reducing DTT) 1. Marker; 2. Human IgG1; 3. ADC standard sample; 4-7: Our ADC sample; 8. Marker

1. Marker; 2. Human IgG1; 3. ADC standard sample; 4-7: Our ADC sample; 8. Marker

2. DAR detect

In this batch, we selected 5.4 equivalent TCEP and 10.8 equivalent VCMMAE groups for DAR detection. The DAR of our sample is almost the same as that of the standard sample, but the integrity of the antibody result is not as good as that of the standard sample.

Figure. The DAR detect of ADC standard sample by LC-MS.

Figure. The DAR detect of our ADC sample by LC-MS. At the beginning of this project, to ensure the antibody can conjugate with payload(MMAE), the dose of TCEP and MMAE is high.
However, we find the dose of TCEP cannot be high, otherwise the damage to the antibody structure is more severe.

3. cytotoxity assay

At next step, we did cytotoxity assay to observe the toxic effect of ADC on 293 cells. At the same time, considering the storage and transportation of ADC, we lyophilized ADC and compared the effect of non-lyophilized on ADC cytotoxicity.

Figure. Our ADC sample deal with 293T.

Obviously, compared with the blank group, ADC had a significant inhibitory effect on the growth of 293 cells. Moreover, lyophilized can reduce the inhibition of ADC on cell growth.

The results show that when the equivalent of reducing agent TCEP is 0.4 and 1.5, the results do not have regularity, which may be caused by the limited opening of disulfide bond and the limited coupling of small molecules. However, when the equivalent of TCEP is 3, the results can match the previous PTM-1-ADC (5.4t5.4v & 5.4t10.8). We also deal 293-ko cell line with PTM-1-ADC, obviously, there is no regular relationship between cell death and ADC dose.

View the Knowledge base of neutralizing antibody (NAb):     - The Landscape of Antibody-drug Conjugate (ADC): Production, Mechanisms of Action (MOA), FDA approved-antibodies, and Functional assay     - What is antibody-drug conjugate (ADC)?     - Antibody-drug conjugate (ADC) in clinical application (Approved/BLA, phaseI/II/III)     - Main elements of antibody-drug conjugate (ADC)： Antibodies and their targets     - Main elements of antibody-drug conjugate (ADC)：Linker (cleavable/non-cleavable, structure and mechanism)     - Main elements of antibody-drug conjugate (ADC)：Toxins/Payloads (Classification and function)     - Toxins/Payloads (Classification and function) of Microtubule destroying drug     - Toxins/Payloads (Classification and function) of DNA damage drugs     - Toxins/Payloads (Classification and function) of Innovative drugs     - Biological coupling technology Chemical based specific in situ antibody modification     - Endogenous coupling of amino acids and Disulfide re bridging strategy     - Glycan coupling     - Site specific biological coupling of engineered antibodies and Enzymatic method     - Biological coupling with engineered unnatural amino acids     - Review for ADC production, quality control and functional assay     - Product data of ADC