Structural study of human antibody fragments with specificity for Mucin-1 antigen
type: CreativeWork
description: MUC1 is overexpressed in many adenocarcinomas, which makes it a potential target for immunotherapy. A large number of MUC1-specific, monoclonal antibodies have been produced but the stability, solubility and the size of the antibody determines their effectiveness in cancer therapy. Smaller antibody fragments are advantageous over full-size antibodies, since small antigen binding molecules can efficiently penetrate solid tumors. As a first step towards the design of a clinically desirable antibody fragment, the crystallization of a human VH fragment has been achieved. The fragment was derived from the human single chain antibody scFvM12, which recognizes the cancer-specific hypoglycosylated MUC1. The human M12-VH domain has been crystallized through limited in-vitro proteolysis of scFvM12 antibody fragment. Crystals of M12-VH domain grew in approximately 12 months up to a size of 180x60x60µm in 100mM MES, pH 6.5, 5mM zinc sulphate, 25% v/v polyethylene glycol 550 monomethylether in the presence of low concentration of subtilisin Carlsberg. The protease addition results in complete degradation of M12-VL domain, linker and purification tags. The crystal belongs to space group C2 with unit cell dimension a = 71.34Å, b = 37.97Å, c = 37.19Å and alpha = gamma = 90°, beta = 109.674°. The solvent content of the crystal was calculated as 35.94%. The structure was solved up to 1.5Å resolution with an Rcryst factor of 15.8%, an Rfree of 19.7% and possesses a good stereochemistry. Dihedral angle values comparison of first and second complementarity-determining region (CDR) of M12-VH domain with an average value, for these two hypervariable regions, shows a significant deviation and therefore, it can be speculated that the M12-VH either suggests the existence of new canonical subclass or a link among the subclasses of canonical main-chain conformation in VH3 family. The natural existence of the stabilizing mutations Glu-H6, Arg-H66 and Ser-H52 confers the intrinsic stability to M…
abstract: MUC1 is overexpressed in many adenocarcinomas, which makes it a potential target for immunotherapy. A large number of MUC1-specific, monoclonal antibodies have been produced but the stability, solubility and the size of the antibody determines their effectiveness in cancer therapy. Smaller antibody fragments are advantageous over full-size antibodies, since small antigen binding molecules can efficiently penetrate solid tumors. As a first step towards the design of a clinically desirable antibody fragment, the crystallization of a human VH fragment has been achieved. The fragment was derived from the human single chain antibody scFvM12, which recognizes the cancer-specific hypoglycosylated MUC1. The human M12-VH domain has been crystallized through limited in-vitro proteolysis of scFvM12 antibody fragment. Crystals of M12-VH domain grew in approximately 12 months up to a size of 180x60x60µm in 100mM MES, pH 6.5, 5mM zinc sulphate, 25% v/v polyethylene glycol 550 monomethylether in the presence of low concentration of subtilisin Carlsberg. The protease addition results in complete degradation of M12-VL domain, linker and purification tags. The crystal belongs to space group C2 with unit cell dimension a = 71.34Å, b = 37.97Å, c = 37.19Å and alpha = gamma = 90°, beta = 109.674°. The solvent content of the crystal was calculated as 35.94%. The structure was solved up to 1.5Å resolution with an Rcryst factor of 15.8%, an Rfree of 19.7% and possesses a good stereochemistry. Dihedral angle values comparison of first and second complementarity-determining region (CDR) of M12-VH domain with an average value, for these two hypervariable regions, shows a significant deviation and therefore, it can be speculated that the M12-VH either suggests the existence of new canonical subclass or a link among the subclasses of canonical main-chain conformation in VH3 family. The natural existence of the stabilizing mutations Glu-H6, Arg-H66 and Ser-H52 confers the intrinsic stability to M…
author: Gaur, Rajneesh Kumar
language: eng
keywords: info:eu-repo/classification/ddc/570, Biowissenschaften, Biologie, Antibody, Mucin-1, Limited Proteolysis, Crystallization
date published: 2004
date modified: 2026-07-30T19:43:19Z
provider: RWTH Aachen University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: Publikationsserver der RWTH Aachen University