Manufacturing Process Development for an Epidermal Growth Factor-Based Cancer Vaccine - By incorporating disposable technologies and an improved purification scheme, scale-up and validation problems w

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Manufacturing Process Development for an Epidermal Growth Factor-Based Cancer Vaccine
By incorporating disposable technologies and an improved purification scheme, scale-up and validation problems were resolved.


BioPharm International Supplements


Evaluating the Equivalence of the Vaccine Preparations

The equivalence of products produced for POC trials and for ASD was evaluated using molecular characterization, biological activity in mice, and clinical data from administration to patients with advanced lung cancer.


Figure 5. Peptide maps of chromatographic fractions corresponding to the conjugated rEGF-rP64k (active moieties) from (A) product made for proof-of-concept trials and (B) product made for advanced stages of development.
To demonstrate equivalence through molecular characterization, the biologically active moieties (rEGF–rP64k conjugated species) of both products (for POC and ASD) were analyzed by peptide mapping, following digestion of the conjugated rEGF-rP64k fractions with the endoprotease Glu-C. Each fraction of the resulting maps was identified and sequenced by mass spectrometry. Figure 5 shows the peptide maps of the rEGF–rP64k conjugated species from A) the product made for POC and B) the product made for ASD. A high degree of similarity is seen in the peptide profiles and their relative amounts, indicating the equivalence of the products in terms of the structure of their biologically active moieties (rEGF–rP64k conjugated species).


Figure 6. Comparison of the immunogenic activity test results obtained after immunizing mice with vaccine preparations for proof-of-concept (POC) trials and for advanced stages of development (ASD). Each bar represents the mean optical density from an ELISA assay performed on eight lots of vaccine. Different sera dilutions were tested (1:100, 1:1,000 and 1:10,000).
Figure 6 shows biological activity using measurements of anti-rEGF antibody response in mice for the POC and the ASD vaccine preparations. No statistically significant differences were observed.


Figure 7. Anti-EGF antibody responses in patients vaccinated with the preparations made for proof-of-concept (POC) trials and advanced stages of development (ASD). Y axis represents the geometric means of anti-EGF antibody titers; bars represent the geometric mean of antibody titers in eight patients vaccinated with the two different products. The line represents the geometric mean of anti-EGF antibody in time for control sera (unvaccinated patients).
The clinical performance of both products was also evaluated in advanced lung cancer patients. Clinical data from eight patients vaccinated with each vaccine preparation were compared (Figure 7). As can be observed, there were no significant differences in the anti-rEGF antibody titers obtained when immunizing patients with the vaccine for POC and with the vaccine for ASD, demonstrating an equivalent immunological effect in humans.

Conclusions

The initial limitations of the manufacturing process used for proof of concept studies of the CIMAvax-EGF vaccine were overcome by a process development strategy that focused on scalability and GMP requirements. The new in-process tests and end-product testing showed acceptable consistency and comparability. The vaccine preparation obtained following the manufacturing improvements showed similar clinical performance compared to the product used during the proof-of-concept trials.

GRYSSELL RODRIGUEZ is the CIMAvax-EGF manufacturing manager, AIRAMA ALBISA is a process development scientist, LISEL VIÑA is the CIMAvax-EGF quality control manager, ARIADNA CUEVAS is the CIMAvax-EGF regulatory affairs manager, BEATRIZ GARCIA is a clinical researcher, AURORA TAMARA GARCIA is the CIMAvax-EGF quality assurance manager, ALEJANDRO PORTILLO is a quality control specialist, LOANY CALVO is a quality control specialist, TANIA CROMBET is the clinical research director, and ERNESTO CHICO is the technical director, all at the Centro de Inmunología Molecular (Center for Molecular Immunology), Havana, Cuba,
, +53-7 2716810. GISELA GONZALEZ is the business development manager at CIMAB SA, Havana, Cuba.


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