Reshaping Human Antibodies: Grafting an Antilysozyme Activity

Verhoeyen, Martine; Milstein, C.; Winter, Greg

doi:10.1126/science.2451287

Cited by 326 publications

(129 citation statements)

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“…It is possible to construct hybrid antibodies by substituting the genetic information for antigen-binding regions of human heavy and light chains with their rodent counterparts of desired antigen specificity (Winter & Milstein, 1991). The whole humanized antibody can then be produced in rat myeloma cells (Reichmann et al, 1988;Verhoeyen et al, 1988;Tempest et al, 1991) or more effectively in Chinese hamster ovary cells (Page & Sydenham, 1991). However, if the Fc portion of a MAb were dispensable for efficient uptake and presentation of antigens by antigen-presenting cells, Fab or Fv fragments could substitute for the whole antibody in SMAA complexes.…”

Section: Discussionmentioning

confidence: 99%

“…An economic approach to construction of SMAA complexes would be to employ a single 'humanized' MAb specific for a small universal peptide tag antigen, used to present any protein to which the tag antigen was attached. Furthermore, the possible use of Fab or Fv fragments for construction of SMAA complexes would enable a largescale production of these fragments using bacterial expression systems (Boulot et al, 1990;Skerra et al, 1991), thus avoiding a laborious and expensive production of whole human antibodies in rat myeloma cell lines (Reichmann et al, 1988;Verhoeyen et al, 1988;Tempest et al, 1991).…”

Section: Introductionmentioning

confidence: 99%

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Construction of solid matrix-antibody-antigen complexes containing simian immunodeficiency virus p27 using tag-specific monoclonal antibody and tag-linked antigen

Hanke

Szawlowski

Randall

1992

Journal of General Virology

141

107

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We have previously shown that immunization with solid matrix-antigen-antibody (SMAA) complexes induces both vigorous humoral and cell-mediated immune responses and have suggested that this method of vaccination may be developed for use in humans, and potentially as a vaccine against AIDS. Here we demonstrate that a small oligopeptide can act as a tag for the construction of SMAA complexes using a tagspecific monoclonal antibody and tag-linked antigens. We show that a 14-amino acid oligopeptide, present in the phospho (P) and V proteins of simian virus 5 (SV5), retains its antigenicity when attached to the C terminus of three 'foreign' proteins [p27 and gp110 of simian immunodeficiency virus (SIV) and glutathione S-transferase] such that these proteins can be incorporated into SMAA complexes using a monoclonal antibody (MAb) that was originally raised against the native SV5 P and V proteins. Mice were immunized with SMAA complexes containing recombinant p27-TAG and MAbs have been isolated that recognized native SIV p27. The significance of these results in terms of the development of SMAA complexes as human vaccines is discussed.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Construction of solid matrix-antibody-antigen complexes containing simian immunodeficiency virus p27 using tag-specific monoclonal antibody and tag-linked antigen

Hanke

Szawlowski

Randall

1992

Journal of General Virology

141

107

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“…The murine V regions can be genetically humanized by the grafting of CDRs into human framework regions. 51 The genetic humanization must take into account the antigen binding properties of the antibody. Some of the framework residues are involved in the antigen binding and some interact with the structures of the CDR loops.…”

Section: Analysis Of Immunogenicity Of Cc49 Scfvsmentioning

confidence: 99%

Effects of humanization and gene shuffling on immunogenicity and antigen binding of anti-tag-72 single-chain Fvs

Pavlínková

Colcher²,

Booth

et al. 2001

Int. J. Cancer

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Key words: antibody engineering; antigen binding; antiidiotypic antibodies; monoclonal antibodies; single-chain antibodiesOne of the major complications in the clinical application of monoclonal antibodies can be the development of a human antimurine antibody (HAMA) response in patients. The HAMA can alter the clearance rate of the administered antibody from serum and can produce allergic reactions in patients. 1,2 The HAMA response can be directed against (i) mouse-specific determinants on the C region of heavy and light chains (antiallotypic Ig), (ii) isotypic determinants on the C region of heavy chains (antiisotypic Ig) and (iii) determinants specific for the administered mouse Ig (antiidiotypic Ig). 3 Many murine monoclonal antibodies (MAbs) have been humanized to overcome the HAMA response in patients. Two methods are being used for the genetic humanization of MAbs: chimerization, in which non-human C regions are replaced by C human regions 4 and complementary determining region (CDR) grafting, in which human CDRs are replaced by murine CDRs on the human framework regions. 5 The humanized MAbs are likely to be less immunogenic; however, murine CDRs are coding for non-human V region sequences and may still elicit an antiidiotypic response. 6 -9 Furthermore, CDR grafting could also generate new immunogenic epitopes. 6,9 The murine MAb CC49 recognizes tumor-associated glycoprotein (TAG-72), which is expressed by most human adenocarcinomas of the pancreas, colon, ovary, prostate, lung and esophagus 10 and is absent in most normal tissues. 11 Radiolabeled CC49 IgG has shown excellent tumor localization in several clinical trials. [12][13][14][15][16] However, one major complication of its clinical use is the development of a HAMA response in patients. 14 The incidence of HAMA response in gastrointestinal and ovarian cancer patients 1 month after exposure to the murine MAb CC49 was 70%. 17 Fifty-four percent of these patients developed an antiidiotypic response, and 12% developed an antiidiotypic response in the absence of antiallotypic and antiisotypic Ig. 17 However, CC49 MAb is less immunogenic than other murine MAbs that have been used in clinical trials. For example, 89%, 100% and 80% of patients receiving the MAbs B72.3, 17 G250 18 and CO17-1A, 19 respectively, developed the HAMA responses.In an attempt to modify the properties of the MAb CC49, genetically engineered CC49 single-chain antibody fragments (scFvs) have been developed. Compared with the corresponding IgG, scFvs exhibited a rapid blood clearance, excellent penetration into the tumor from the vasculature and higher tumor/normal tissue ratios in animal models. 20 -31 We have undertaken new studies to analyze the immunogenicity of CC49 scFvs using human sera obtained from patients with colon cancer treated by CC49 IgG in clinical trials. Three different CC49 scFv constructs (Fig. 1) were compared. The first construct was comprised of mouse CC49 variable light (V L ) and mouse CC49 variable heavy (V H ) regions connected by a 205C linker (m/m scF...

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“…A recent development has been the recombinant production of Fv fragments. These are the smallest possible intact fragments of the antibody molecule that can bind antigen in the appropriate configuration (Verhoeyen et al, 1988 A second possibility to increase tumour uptake of antibody is to give a much higher amount of antibody as long as this is relatively non-toxic to the host. It would be advantageous if the cytotoxic moeity or imaging isotope could then be delivered at a time when the tumour to nontumour ratios are optimal and at a time when there was no significant circulating amount of antibody.…”

Section: Human Milk Fat Globule Membranementioning

confidence: 99%