Rapid expression cloning of human immunoglobulin Fab fragments for the analysis of antigen specificity of B cell lymphomas and anti-idiotype lymphoma vaccination

Idiotype-pulsed dendritic cell vaccination for B-cell lymphoma: clinical and immune responses in 35 patients

“…6,22,39 Moreover, novel genetic approaches are now yielding recombinant forms of Id and will continue to make large-scale production of Id vaccines more practical. [52][53][54][55][56] These technologies offer new sources of Id for loading into DCs. In the future, as additional and possibly shared NHL-associated tumor antigens are described, these too will be candidates for evaluation using DC-based vaccine strategies.…”

Phase I Trial of a Novel Intradermal Idiotype Vaccine in Patients with Advanced B-Cell Lymphoma: Specific Immune Responses Despite Profound Immunosuppression

“…The expression strategy as a recombinant Fab fragment in E. coli (20) has a success rate of f90% and permits substantially reduced manufacturing times. Although lack of glycosylation of these bacterially expressed vaccines could potentially affect their efficacy, experimental evidence suggests that immunization with unglycosylated antigen may not impair but rather enhance cellular immune responses against naturally glycosylated peptides (30,31).…”

“…GMPgrade individual idiotype vaccines were produced commercially (CellGenix GmbH, Freiburg, Germany) by anchored reverse transcription-PCR cloning of the idiotype heavy (H) and light (L) chain expressed by the lymphoma. Lymphoma-derived variable immunoglobulin segments were inserted into pFab.gn or pFab.gE, and recombinant idiotype protein was expressed in E. coli as a recombinant IgG1 Fab fragment (20). Fab protein was purified by affinity chromatography and gel filtration.…”

“…To ease manufacturing of individual idiotype vaccines and to shorten production times, we have adapted a bacterial expression system for recombinant immunoglobulin Fab fragments to the production of recombinant idiotype vaccines in Escherichia coli (19,20). Stimulation of peripheral blood T cells from lymphoma patients with such recombinant, lymphoma-derived Fab fragment resulted in specific, MHC class I-restricted cytotoxic activity against autologous, idiotype-expressing target cells (21).…”

Anti-idiotype vaccines for human follicular lymphoma

“…[46][47][48][49] However, these results have yet to be translated into potentially reliable clinical applications. Positive evidence of potential efficacy of this approach in humans will be required before the various intriguing methods that have been developed for rapid production of genetic Id vaccines, such as their expression in tobacco plants 50 or in E. coli, 51 can have a chance to pay off. In this regard, a major trial has just started at Southampton University to test one such vaccine in humans.…”