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The Bacillus Calmette-Gu\u00e9rin (BCG) vaccine, developed in 1921, remains the primary defense against tuberculosis but requires modernized production methods. The recombinant BCG-pertussis strain shows potential in providing dual protection against tuberculosis and whooping cough, especially for vulnerable newborns, and enhanced efficacy against bladder cancer. Implementing submerged cultivation techniques for rBCG-pertussis production can offer increased productivity and standardization. Methods: This study explores a fed-batch cultivation strategy with pH-stat control to feed L-glutamic acid through the acid pump into 1 L bioreactor. Three pH values were evaluated for fed-batch and a simple batch without pH control was done for comparison. The viable cell concentration was compared before and after freeze-drying samples harvested during the cultures. Results: L-glutamic acid was identified as the preferred substrate for rBCG-pertussis. While the fed-batch strategy did not enhance the maximum specific growth rate compared to simple batch cultivation, it did improve the specific growth rate after day 4 in the pH 7.4-controlled fed-batch cultures, thereby reducing the cultivation time. Fed-batch cultures controlled at three pH levels exhibited lower optical density than the simple batch, although the viable cell counts were similar. Notably, samples harvested after day 8 from the simple batch cultures showed a reduction in CFU\/mL after freeze-drying, whereas all fed-batch samples exhibited high recovery of viable cell counts post lyophilization. Conclusions: The additional glutamate supplied to the fed-batch cultures may have protected the cells during the lyophilization process.<\/jats:p>","DOI":"10.3390\/pharmaceutics16111433","type":"journal-article","created":{"date-parts":[[2024,11,11]],"date-time":"2024-11-11T03:52:07Z","timestamp":1731297127000},"page":"1433","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Comparison Between Simple Batch and Fed-Batch Bioreactor Cultivation of Recombinant BCG"],"prefix":"10.3390","volume":"16","author":[{"given":"Sarah","family":"Mendes","sequence":"first","affiliation":[{"name":"Instituto Butantan, S\u00e3o Paulo 05503-900, Brazil"},{"name":"Interunits Graduate Program in Biotechnology (PPIB), University of S\u00e3o Paulo, S\u00e3o Paulo 05508-220, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4510-1393","authenticated-orcid":false,"given":"Maria C. P.","family":"Gon\u00e7alves","sequence":"additional","affiliation":[{"name":"Instituto Butantan, S\u00e3o Paulo 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-9867-8505","authenticated-orcid":false,"given":"Vitoria A. P.","family":"Aiex","sequence":"additional","affiliation":[{"name":"Instituto Butantan, S\u00e3o Paulo 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryh\u00e1ra D.","family":"Batista","sequence":"additional","affiliation":[{"name":"Instituto Butantan, S\u00e3o Paulo 05503-900, Brazil"},{"name":"Interunits Graduate Program in Biotechnology (PPIB), University of S\u00e3o Paulo, S\u00e3o Paulo 05508-220, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-0700-1997","authenticated-orcid":false,"given":"Patr\u00edcia","family":"Zorzete","sequence":"additional","affiliation":[{"name":"Instituto Butantan, S\u00e3o Paulo 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0156-1312","authenticated-orcid":false,"given":"Luciana C. C.","family":"Leite","sequence":"additional","affiliation":[{"name":"Instituto Butantan, S\u00e3o Paulo 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0980-8116","authenticated-orcid":false,"given":"Viviane M.","family":"Gon\u00e7alves","sequence":"additional","affiliation":[{"name":"Instituto Butantan, S\u00e3o Paulo 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,11,11]]},"reference":[{"key":"ref_1","unstructured":"(2024, July 21). WHO: Global Tuberculosis Report: 2023. Available online: https:\/\/www.who.int\/teams\/global-tuberculosis-programme\/tb-reports\/global-tuberculosis-report-2023."},{"key":"ref_2","unstructured":"(2024, July 21). WHO: Global Tuberculosis Report: 2019. 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