Antibody Fragmentation Services
Bio-Synthesis offers antibody fragmentation service to customers by selectively cleave immunoglobulin into Fab, Fab', F(ab')2 and Fc fragments using papain, pepsin or ficin to avoid protease contamination and provides precise control of the digestion process. Our antibody fragmentation service includes:
- Free consultation of antibody fragmentation
- Antibody preparation and fragmentation
- Concentration and rebuffering
- OD280 determination
- FPLC-based purification control
- Determination of total yield
- Documentation
| Available Services |
Source |
|
| Fc Fragmentation |
Human or mouse IgG antibody |
| Fab Fragmentation |
Human or mouse IgG antibody |
| F(ab')2 Fragmentation |
Whole human and other igG antibody |
| Mouse IgG1 Fab and F(ab')2 |
Mouse IgG1 |
| IgM Fragmentation (Fab or IgG) |
Whole IgM antibody |
|
|
Ordering:
Quality Control:
SDS-PAGE gel electrophoresis (optional): Analysis of antibody fragments by SDS PAGE gel electrophoresis
(Includes 2 mini gels with 1 standard each and up to 8 samples per gel and a copy of COA)
To obtain further information,
contact us. Our technical staff is also available to assist you in determining the type of production system that best meets your needs
Enzymes used for Fragmentation
Fragmentation of IgG and IgM from a variety of species
Papain
Papain is a nonspecific, thiol-endopeptidase that has a sulfhydryl group in the active site, which
must be in the reduced form for activity. Papain has been shown to produce heterogeneous fragments from IgM. Oligosaccharides in the hinge region of IgM interfere with papain digestion, causing a cleavage shift of 3-5 amino acids in either direction.
Pepsin
Pepsin is a nonspecific endopeptidase that is active only at an acid pH, and it is irreversibly denatured at neutral or alkaline pH. It is possible to produce F(ab')2, Fab and Fv fragments using pepsin to digest IgM.
Trypsin
Trypsin is a serine protease that reacts optimally at pH 8.0. In general, increasing the enzyme/substrate ratio and/or temperature will increase the rate of digestion. Trypsin can generate F(ab')2, Fab, "IgG" - type and Fc5µ fragments from IgM. Trypsin digestion of several species of IgM was studied using trypsin with and without urea pretreatment. 26 Urea alters the susceptibility of the domains to digestion and produces different fragments than those digested in aqueous buffer. Many other procedures have been developed to digest IgM using trypsin.
2-Mercaptoethylamine
Mild reduction can be achieved using 2-mercaptoethylamine (2-MEA), Hydrochloride (HCl). Reduction
will vary among IgM species, but an "IgG" - type and/or reduced IgG ("rIgG") should be formed in
varied proportions, depending upon reduction time and/or temperature (Figure 11).36 Fragmentation of mouse IgM also produces an inverted "IgG"-type fragment.