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This study focuses on two surface proteins from two <jats:italic>Borrelia<\/jats:italic> subspecies involved in these diseases: CspZ, expressed by <jats:italic>Borrelia burgdorferi<\/jats:italic> sensu stricto (also named BbCRASP-2 for complement regulator-acquiring surface protein 2), and the factor H binding A (FhbA), expressed by <jats:italic>Borrelia hermsii.<\/jats:italic> Numerous subspecies of <jats:italic>Borrelia<\/jats:italic>, including these latter, are able to evade the immune defenses of a variety of potential vertebrate hosts in a number of ways. In this context, previous data suggested that both surface proteins play a role in the immune evasion of both <jats:italic>Borrelia<\/jats:italic> subspecies by interacting with key regulators of the alternative pathway of the human complement system, factor H (FH) and FH-like protein 1 (FHL-1). The recombinant proteins, CspZ and FhbA, were expressed in <jats:italic>Escherichia coli<\/jats:italic> and purified by one-step metal-affinity chromatography, with yields of 15 and 20\u00a0mg or pure protein for 1\u00a0L of cultured bacteria, respectively. The purity was evaluated by SDS-PAGE and HPLC and is close to about 95%. The mass of CspZ and FhbA was checked by mass spectrometry (MS). Proper folding of CspZ and FhbA was confirmed by circular dichroism (CD), and their biological activity, namely their interaction with purified FH from human serum (recombinant FH<jats:sub>15-20<\/jats:sub>\u00a0and recombinant FHL-1), was characterized by SPR. Such a study provides the basis for the biochemical characterization of the studied proteins and their biomolecular interactions which is a necessary prerequisite for the development of new approaches to improve the current diagnosis of LB and TBRF.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Key points<\/jats:title>\n            <jats:p>\u2022<jats:italic> DLS, CD, SEC-MALS, NMR, HPLC, and MS are tools for protein quality assessment<\/jats:italic>\n            <\/jats:p>\n            <jats:p>\u2022<jats:italic> Borrelia spp. possesses immune evasion mechanisms, including human host complement<\/jats:italic>\n            <\/jats:p>\n            <jats:p>\u2022<jats:italic> CspZ and FhbA interact with high affinity (pM to nM) to human FH and rFHL-1<\/jats:italic>\n            <\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Graphical Abstract<\/jats:title>\n          <\/jats:sec>","DOI":"10.1007\/s00253-024-13195-2","type":"journal-article","created":{"date-parts":[[2024,7,23]],"date-time":"2024-07-23T07:01:47Z","timestamp":1721718107000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Production, purification, and quality assessment of borrelial proteins CspZ from Borrelia burgdorferi and FhbA from Borrelia hermsii"],"prefix":"10.1007","volume":"108","author":[{"given":"Micka\u00ebl","family":"Gu\u00e9rin","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maryl\u00e8ne","family":"Vandevenne","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alain","family":"Brans","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e9","family":"Matagne","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rodrigue","family":"Marquant","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elise","family":"Prost","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"St\u00e9phane","family":"Octave","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B\u00e9rang\u00e8re","family":"Avalle","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irene","family":"Maffucci","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5475-1358","authenticated-orcid":false,"given":"S\u00e9verine","family":"Padiolleau-Lef\u00e8vre","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,7,23]]},"reference":[{"key":"13195_CR1","doi-asserted-by":"publisher","first-page":"705","DOI":"10.1111\/j.1651-2227.1981.tb05772.x","volume":"70","author":"M Adinolfi","year":"1981","unstructured":"Adinolfi M, Dobson NC, Bradwell AR (1981) Synthesis of two components of human complement, \u03b21 h and C3bINA, during fetal life. 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