膜杰作
Star Staining
组分（Materials Provided）
| ID | Components | Size |
| ADC-P008-C01 | 20 x Reaction buffer | 50ul |
| ADC-P008-C02 | Cofactor A | 50ul |
| ADC-P008-C03 | Cofactor B | 50ul |
| ADC-P008-C04 | Substrate A | 25ul |
| ADC-P008-C05 | Substrate B | 25ul |
| ADC-P008-C06 | Enzyme A | 25ul |
| ADC-P008-C07 | Enzyme B | 25ul |
| ADC-P008-C08 | Enzyme C | 20ul |
| ID | Components | Size |
| ADC-P008-1-C01 | Binding buffer | 15ml |
| ADC-P008-1-C02 | Elution buffer | 1.5ml |
| ADC-P008-1-C03 | Neutralization buffer | 0.5ml |
| ADC-P008-1-C04 | 250 mM Tris-HCl buffer (pH7.5) | 1.5ml |
| ADC-P008-1-C05 | 10×PBS (pH7.2- 7.4) | 6ml |
| ADC-P008-1-C06 | Desalting column | 2per |
| ADC-P008-1-C07 | Concentrator tube | 2per |
| ADC-P008-1-C08 | ProteinA Resin | 100ul*2 |
背景（Background）
Antibody-drug conjugates (ADC) is a type of fast growing anticancer drug. Monoclonal antibody is conjugated to the cytotoxic payload via a chemical linker that directed toward a target antigen expressed on the cancer cell surface, reducing systemic exposure and therefore toxicity. AGLink® ADC conjugation kit is designed to conjugate mAb with Biotin though site-specific enzymatic reaction.
应用说明（Application）
AGLink® ADC Conjugation Kit (Biotin, DAR2&4, 1mg) is designed to conjugate 1mg mAb with Biotin through site-specific enzymatic reaction.
It is for research use only.
优势特色（Features）
- Site-specific conjugation through Fc-glycan with high conjugation efficiency
- Uniform DAR value and highly homogeneous ADC products
- No need of amino acid sequence engineering
- Easy to use: One-pot & One-step
- Two options: DAR2 & DAR4 ADC product
存储（Storage）
BoxA: The unopened kit is stable for 12 months from the date of manufacture if stored under temperature ≤ -20°C. The opened kit should be stored according to components table. BoxB: The unopened kit is stable for 12 months from the date of manufacture if stored under 4°C. The opened kit should be stored according to components table.
原理-Assay Principles
Almost all monoclonal antibodies are glycosylated at (or around) Asn-297 of the Fc domain. While the glycans have different isoforms, the types that typically dominate are G0F, G1F&G2F (＞90%). The A&Glink® Antibody Conjugation Kit utilizes the YTConju™ platform, a glycan remodeling strategy developed by Glyco-therapy Biotechnology Co., Ltd. for the construction of site-specific antibody conjugates.
The A&Glink® DAR2&4 Antibody Conjugation Kit integrates the procedure for constructing DAR2 conjugates and DAR4 conjugates in one kit, providing flexibility in DAR design.
To construct the DAR4 conjugates, galactose is first added to the terminal GlcNAc catalyzed by β-1,4-galactosyltransferase (GT) to form LacNAc. Then, α-1,3-fucosyltransferase (FT) recognizes LacNAc and transfers the Fuc-Payload to the GlcNAc of the LacNAc to form a conjugate with a theoretical DAR of 4.
To construct the DAR2 conjugates, glycans are first removed from the core GlcNAc catalyzed by endoglycosidase. Galactose is then added to the GlcNAc catalyzed by β-1,4-galactosyltransferase (GT) to form core LacNAc. Finally, α-1,3-fucosyltransferase (FT) recognizes LacNAc and transfers the Fuc-Payload to the GlcNAc of the LacNAc to form a conjugate with a theoretical DAR of 2.
The DAR4 and DAR2 conjugation procedures share the same FT, GT and GDP-Fuc-Payload. The main difference between these two procedures is the addition of an endoglycosidase.
These procedures are performed in a “one-pot” manner with only one purification step, which is highly convenient and efficient. Antibody conjugates produced with this kit generally exhibit high homogeneity, high stability and high hydrophilicity.
典型数据-Typical Data Please refer to DS document for the assay protocol.
ADC DAR value assessment by LC-TOF/MS
1.Analyst TF 1.8.1 Software was used to collect LC-TOF/MS data of samples in positive ion mode
2.PeakView was used for data processing and BioToolKit was used for deconvolution analysis.
3.DAR values are calculated using the following formula:
DAR4 =(920.12*4+77.42*3)/(920.12+77.42)=3.92
ADC DAR value assessment by LC-TOF/MS
1.Analyst TF 1.8.1 Software was used to collect LC-TOF/MS data of samples in positive ion mode
2.PeakView was used for data processing and BioToolKit was used for deconvolution analysis.
3.DAR values are calculated using the following formula:
DAR4 =（1138.27*2）/1138.27=2.00
Statement of Rights
AGLink products are co-developed by ACROBiosystems Co., Ltd. and Glyco-therapy Biotechnology Co., Ltd. based on Glyco-therapy's conjugation platform (YTConju™) and are not authorized for commercial use. The sale and use of AGLink products doesn't represent any forms of license or authorization of the attached technologies for developing any diagnostic, therapeutic or prophylactic products for commercial purposes.
Note of application
AGLink products are intended for research use only by the Buyer or its users and can not be used for any other purposes, including, but not limited to, use for any clinical diagnostic type use, in vitro or in vivo therapeutic use, prophylactic use, any type of consumption or application to human or animals.
