![]() ![]() Note that at this point only the glycosylation of one subunit can be defined. Now you can enter the position (298) and the subunit (1) into the appropriate box. As only the asparagine at the position 298 on the HC is glycosylated, select “Partly” option. Figure 3: Selection of modification.Ī new window is popped up where you can specify the glycosylation (figure 4). Choose “N-Glycosylation” within the “Modifications” tab and press the button “Add Modification” (figure 3). Figure 2: Entered amino acid sequence of two HC and two LC. SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTĬopy the headline and the amino acid sequence of the LC in the same matter into the third and the fourth text area (figure 2). RFSGSGSGTDFTLTINSLEAEDAAAYYCHQSSSLPFTFGPGTKVDIKRTVAAPSVFIFPP GNVFSCSVMHEALHNHYTQKSLSLSPGK Light chain: >8836_L|canakinumab|Homo sapiens||L-KAPPA (V-KAPPA(1-107)+C-KAPPA(108-214))|||||||214||||MW 23357.9|MW 23357.9|ĮIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLHWYQQKPDQSPKLLIKYASQSFSGVPS KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMT VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST ![]() YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPS TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL QVQLVESGGGVVQPGRSLRLSCAASGFTFSVYGMNWVRQAPGKGLEWVAIIWYDGDNQYYĪDSVKGRFTISRDNSKNTLYLQMNGLRAEDTAVYYCARDLRTGPFDYWGQGTLVTVSSAS Then copy the amino acid sequence of the HC on a new line below the headline in the first and the second text area. ![]() The headlines are optional, but are helpful for the orientation in the results. Figure 1: Add multiple subunits.Įnter the headline (starting with a “>”) into the first and the second text area followed by a “return”. Therefore add three subunits to a total of four subunits by clicking three times the “add” button (figure 1). The IgG is composed of four subunits – two identical heavy chains (HC) and two identical light chains (LC). For this tutorial the sequence of Canakinumab, a recombinant human anti-human-IL-1β IgG, from DrugBank.ca was used. How-toįirst of all go to the Protein Tool of Prot pi and enter the amino acid sequence of your protein. Even though, in most cases, immunoglobulin G (IgG) is additionally modified, no further posttranslational modifications were described in this article. This short guide deals with how to add two complex-type N-Glycosylation G1 with a sialic acid (N-acetylneuraminic acid) to the amino acid sequence of the heavy chains (HC) of a monoclonal antibody. Therefore Prot pi provides a tool to draw glycans as a posttranslational modification of proteins. Glycosylations should not be neglected for the correct calculation of the molecular mass, the isoelectric point and the mass-specific UV absorption coefficient. ![]()
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