Surface Engineering of the RNA Coliphage Q? to Display Plasmodium Falciparum Derived Asexual Blood Stage Antigens UB05 and Merozoite Surface Protein 3

Waffo, Alain Bopda; Lissom, Abel; Ouambo, Herve F; Ngu, Loveline N.; Ngoh, Apeh A.; Sanders, Carrie; Bawage, Swapnil; Tchadji, Colince J.; Assob, Jules N.C.; Okoli, Arinze S.; Takoua, Doline; Ambada, Georgia; Priso, Ghislain Donald Njambe; Garcia, Rosario; Gutiérrez, Anna; Ndjonka, Dieudonné; Chukwuma, George O; Okeke, Malachy Ifeanyi; Megnekou, Rosette; Esimone, Charles Okechukwu; Achidi, Eric A.; Mbacham, Wilfred N.; Kaptué, Lazare; Park, Chae Gyu; Nchinda, Godwin

doi:10.4172/2327-5073.1000314

Cited by 8 publications

(23 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The entire cDNA of the Qβ phage was cloned into several plasmids and used to produce phage without lysis of the host cell. Together, with the promising new advances in evolution and the uncover on the Qβ genome, we suggest that more factors than A 2 mediating cell lysis are imminent [ 55 , 56 , 57 , 58 , 77 , 78 , 79 , 80 ].…”

Section: Uniqueness Of the Biology Coliphage Qβmentioning

confidence: 86%

See 1 more Smart Citation

Uniqueness of RNA Coliphage Qβ Display System in Directed Evolutionary Biotechnology

Nchinda

Al-Atoom

Coats

et al. 2021

Viruses

View full text Add to dashboard Cite

Phage display technology involves the surface genetic engineering of phages to expose desirable proteins or peptides whose gene sequences are packaged within phage genomes, thereby rendering direct linkage between genotype with phenotype feasible. This has resulted in phage display systems becoming invaluable components of directed evolutionary biotechnology. The M13 is a DNA phage display system which dominates this technology and usually involves selected proteins or peptides being displayed through surface engineering of its minor coat proteins. The displayed protein or peptide’s functionality is often highly reduced due to harsh treatment of M13 variants. Recently, we developed a novel phage display system using the coliphage Qβ as a nano-biotechnology platform. The coliphage Qβ is an RNA phage belonging to the family of Leviviridae, a long investigated virus. Qβ phages exist as a quasispecies and possess features making them comparatively more suitable and unique for directed evolutionary biotechnology. As a quasispecies, Qβ benefits from the promiscuity of its RNA dependent RNA polymerase replicase, which lacks proofreading activity, and thereby permits rapid variant generation, mutation, and adaptation. The minor coat protein of Qβ is the readthrough protein, A1. It shares the same initiation codon with the major coat protein and is produced each time the ribosome translates the UGA stop codon of the major coat protein with the of misincorporation of tryptophan. This misincorporation occurs at a low level (1/15). Per convention and definition, A1 is the target for display technology, as this minor coat protein does not play a role in initiating the life cycle of Qβ phage like the pIII of M13. The maturation protein A2 of Qβ initiates the life cycle by binding to the pilus of the F+ host bacteria. The extension of the A1 protein with a foreign peptide probe recognizes and binds to the target freely, while the A2 initiates the infection. This avoids any disturbance of the complex and the necessity for acidic elution and neutralization prior to infection. The combined use of both the A1 and A2 proteins of Qβ in this display system allows for novel bio-panning, in vitro maturation, and evolution. Additionally, methods for large library size construction have been improved with our directed evolutionary phage display system. This novel phage display technology allows 12 copies of a specific desired peptide to be displayed on the exterior surface of Qβ in uniform distribution at the corners of the phage icosahedron. Through the recently optimized subtractive bio-panning strategy, fusion probes containing up to 80 amino acids altogether with linkers, can be displayed for target selection. Thus, combined uniqueness of its genome, structure, and proteins make the Qβ phage a desirable suitable innovation applicable in affinity maturation and directed evolutionary biotechnology. The evolutionary adaptability of the Qβ phage display strategy is still in its infancy. However, it has the potential to evolve functional domains of the desirable proteins, glycoproteins, and lipoproteins, rendering them superior to their natural counterparts.

show abstract

Section: Uniqueness Of the Biology Coliphage Qβmentioning

confidence: 86%

“…The phage Qβ would not be an exception to the rules of RNA viruses, but would be a powerful tool for an IVE study instead. In fact, RNA Qβ phage could be used as a toolkit that would employ and exploit its unique features in the directed evolutionary process [ 54 , 55 , 56 , 57 , 58 ].…”

Section: Rna Evolutionmentioning

confidence: 99%

Uniqueness of RNA Coliphage Qβ Display System in Directed Evolutionary Biotechnology

Nchinda

Al-Atoom

Coats

et al. 2021

Viruses

View full text Add to dashboard Cite

show abstract

“…The antigens consisted of recombinant Qβ displaying Plasmodium falciparum 3D7 strain sequence derived C-terminal part of MSP3 (QβMSP3) and UB05 (QβUB05) generated in our group as previously described [32].…”

Section: Study Antigensmentioning

confidence: 99%

“…The plasma levels of antibodies specific to the malaria antigens QβUB05 and QβMSP3 were determined using the enzyme linked immunosrbent assay, (ELISA) as previously described in our group [16,29,32]. Briefly high binding ELISA plates were coated with 10 7 particles/well of each recombinant phage and incubated overnight at 4°C.…”

Section: Determination Of Igg and Igg Subclasses Antibody Responses S...mentioning

confidence: 99%

Comparative Analysis of Igg Responses to Recombinant Qβ Phage Displayed MSP3 and UB05 in Dual HIV-Malaria Infected Adults Living in areas differing in Malaria Transmission Intensities

2019

JCEI

View full text Add to dashboard Cite

Immunoglobulin G (IgG) specific responses against Plasmodium falciparum merozoite antigens such as the merozoite surface protein 3 (MSP3) and UB05 are known to play critical roles in parasiteamia control and protection from symptomatic illness. However when there is intense perennial malaria transmission coupled with concurrent infection with the human immunodeficiency virus type 1 (HIV), knowledge of IgG antibody response profiles is limited. In this study we assessed the impact of dual HIV-Malaria infections on IgG subclass responses to MSP3 (QβMSP3) and UB05 (QβUB05) in individuals living in two areas of Cameroon differing in malaria transmission intensity. We observed differences in antigen specific IgG and IgG subclass responses which were dependent upon the antigen type, malaria transmission intensity, HIV infection, malaria infection and dual HIV-malaria infections. Individuals living in high malaria transmission areas irrespective of HIV or malaria status had significantly higher IgG responses to both antigens (P=0.0001 for QβMSP3, P=0.0001 for QβUB05) than their counterpart from low transmission areas. When dual HIV-Malaria infection is considered significantly higher QβMSP3 specific IgG1 (P=0.0001) and IgG3 (P=0.04) responses in double negative individuals was associated with protection against malaria in low transmission areas. Increased QβUB05 specific IgG1 responses (P=0.0001) in double negative individuals were associated with protection in high transmission areas in contrast to significantly higher IgG3 responses to QβUB05 (P=0.0001) which were more relevant to protection in low malaria transmission areas in the same population. These findings imply that QβMSP3 might not be suitable as a standalone vaccine in areas differing in transmission intensity. However, antigenicity of UB05 most likely predicts immunity in both low and high transmission areas and could be used either alone or in combination with other antigens for vaccine studies in areas differing in transmission intensities. Understanding immune responses to QβUB05 and QβMSP3 could thus enable the development of efficacious vaccines or commensurate immunotherapeutic strategies suitable for areas differing in malaria transmission intensity.

show abstract

“…During this study we have mapped immunological and host receptor-binding motifs of the SARS-CoV S protein following display upon the RNA coliphage display Qubevirus (Qβ) platform. This platform has recently been shown to expose several functional peptides without compromising the recombinant phage viability (36)(37)(38)(39)(40). Like CoVs, Qβ is a single stranded positive-sense RNA bacteriophage (41).…”

Section: Introductionmentioning

confidence: 99%

Mapping immunological and host receptor binding determinants of SARS-CoV spike protein utilizing the Qubevirus platform

Sanders¹,

Dzelamonyuy²,

Ntemafack³

et al. 2023

Preprint

View full text Add to dashboard Cite

The motifs involved in tropism and immunological interactions of SARS-CoV spike (S) protein were investigated utilizing the Qubevirus platform. We showed that separately, 14 overlapping peptide fragments representing the S protein (F1-14 of 100 residues each) could be inserted into the C-terminus of A1 on recombinant Qubevirus without affecting its viability. Additionally, recombinant phage expression resulted in the surface exposure of different engineered fragments in an accessible manner. The F6 from S425-525, was found to contain the binding determinant of the recombinant human angiotensin converting enzyme 2 (rhACE2), with the shortest active binding motif situated between residues S437-492. Upstream, another fragment, F7, containing an overlapping portion of F6 would not bind to rhACE2, confirming not just only that residues were linear but equally also the appropriate structural orientation of F6 upon the Qubevirus. The F6 (S441-460) and other inserts, including F7/F8 (S601-620) and F10 (S781-800), were demonstrated to contain important immunological determinants through recognition and binding of S protein specific (anti-S) antibodies. An engineered chimeric insert bearing the fusion of all three anti-S reactive epitopes, improved substantially the recognition and binding to their cognate antibodies. These results provide insights into humoral immune relevant epitopes and tropism characteristics of the S protein with implications for the development of subunit vaccines or other biologics against SARS-CoV.

show abstract

Surface Engineering of the RNA Coliphage Q? to Display Plasmodium Falciparum Derived Asexual Blood Stage Antigens UB05 and Merozoite Surface Protein 3

Cited by 8 publications

References 34 publications

Uniqueness of RNA Coliphage Qβ Display System in Directed Evolutionary Biotechnology

Uniqueness of RNA Coliphage Qβ Display System in Directed Evolutionary Biotechnology

Comparative Analysis of Igg Responses to Recombinant Qβ Phage Displayed MSP3 and UB05 in Dual HIV-Malaria Infected Adults Living in areas differing in Malaria Transmission Intensities

Mapping immunological and host receptor binding determinants of SARS-CoV spike protein utilizing the Qubevirus platform

Contact Info

Product

Resources

About