title: Efficient recovery of recombinant proteins from cereal endosperm is affected by interaction with endogenous storage proteins.
abstract: Cereal seeds are versatile platforms for the production of recombinant proteins because they provide a stable environment for protein accumulation. Endogenous seed storage proteins, however, include several prolamin-type polypeptides that aggregate and crosslink via intermolecular disulfide bridges, which could potentially interact with multimeric recombinant proteins such as antibodies, which assemble in the same manner. We investigated this possibility by sequentially extracting a human antibody expressed in maize endosperm, followed by precipitation in vitro with zein. We provide evidence that a significant proportion of the antibody pool interacts with zein and therefore cannot be extracted using non-reducing buffers. Immunolocalization experiments demonstrated that antibodies targeted for secretion were instead retained within zein bodies because of such covalent interactions. Our findings suggest that the production of soluble recombinant antibodies in maize could be enhanced by eliminating or minimizing interactions with endogenous storage proteins.
authors: Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Austria.
authors: Jenny
authors: J
authors: Peters
authors: Maite
authors: M
authors: Sabalza
authors: Koreen
authors: K
authors: Ramessar
authors: Paul
authors: P
authors: Christou
authors: Teresa
authors: T
authors: Capell
authors: Eva
authors: E
authors: Stöger
authors: Elsa
authors: E
authors: Arcalís
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000080908:Broadly Neutralizing Antibodies; D002523:Edible Grain / Q000235:genetics* / Q000378:metabolism; D056625:Endosperm / Q000235:genetics / Q000378:metabolism; D015483:HIV Antibodies; D006801:Humans; D058634:Molecular Farming; D010940:Plant Proteins / Q000378:metabolism; D046549:Plantibodies / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification*; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D055314:Seed Storage Proteins / Q000737:chemistry* / Q000378:metabolism; D012639:Seeds / Q000254:growth & development; D003313:Zea mays / Q000196:embryology* / Q000235:genetics / Q000378:metabolism; D015028:Zein / Q000737:chemistry / Q000378:metabolism
chemical_list: C529157:2G12 monoclonal antibody; D000911:Antibodies, Monoclonal; D000080908:Broadly Neutralizing Antibodies; D015483:HIV Antibodies; D010940:Plant Proteins; D046549:Plantibodies; D011994:Recombinant Proteins; D055314:Seed Storage Proteins; D015028:Zein
keywords: Maize endosperm; Molecular farming; Plantibodies; Protein trafficking