Construction and characterization of a humanized, internalizing, B-cell (CD22)-specific, leukemia/lymphoma antibody, LL2

Mol Immunol. 1995 Dec;32(17-18):1413-27. doi: 10.1016/0161-5890(95)00080-1.


The murine monoclonal antibody, LL2, is a B-cell (CD22)-specific IgG2a which has been demonstrated to be clinically significant in the radioimmunodetection of non-Hodgkin's B-cell lymphoma. The antibody carries a variable region-appended glycosylation site in the light chain and is rapidly internalized upon binding to Raji target cells. Humanization of LL2 was carried out in order to develop LL2 as a diagnostic and immunotherapeutic suitable for repeated administration. Based on the extent of sequence homology, and with the aid of computer modeling, we selected the EU framework regions (FR) 1, 2 and 3, and the NEWM FR4 as the scaffold for grafting the heavy chain complementarity determining regions (CDRs), and REI FRs for that of light chains. The light chain glycosylation site, however, was not included. Construction of the CDR-grafted variable regions was accomplished by a rapid and simplified method that involved long DNA oligonucleotide synthesis and the polymerase chain reaction (PCR). The humanized LL2 (hLL2), lacking light chain variable region glycosylation, exhibited immunoreactivities that were comparable to that of chimeric LL2 (cLL2), which was shown previously to have antigen-binding properties similar to its murine counterpart, suggesting that the VK-appended oligosaccharides found in mLL2 are not necessary for antigen binding. Moreover, the hLL2 retained its ability to be internalized into Raji cells at a rate similar to its murine and chimeric counterparts.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Neoplasm / biosynthesis*
  • Antibodies, Neoplasm / chemistry
  • Antibodies, Neoplasm / genetics
  • Antibody Specificity / genetics
  • Antigens, CD / genetics*
  • Antigens, Differentiation, B-Lymphocyte / genetics*
  • Base Sequence
  • Cell Adhesion Molecules*
  • DNA Primers
  • Humans
  • Lectins*
  • Leukemia, B-Cell / genetics
  • Leukemia, B-Cell / immunology
  • Lymphoma, B-Cell / genetics*
  • Lymphoma, B-Cell / immunology*
  • Lymphoma, B-Cell / metabolism
  • Mice
  • Molecular Sequence Data
  • Oligonucleotides / chemical synthesis
  • Polymerase Chain Reaction
  • Protein Engineering
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / chemistry
  • Sialic Acid Binding Ig-like Lectin 2
  • Tumor Cells, Cultured


  • Antibodies, Neoplasm
  • Antigens, CD
  • Antigens, Differentiation, B-Lymphocyte
  • CD22 protein, human
  • Cd22 protein, mouse
  • Cell Adhesion Molecules
  • DNA Primers
  • Lectins
  • Oligonucleotides
  • Recombinant Fusion Proteins
  • Sialic Acid Binding Ig-like Lectin 2

Associated data

  • PDB/1REI
  • PDB/3FAB