Subcutaneous delivery of an antibody against SARS-CoV-2 from a supramolecular hydrogel depot
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Abstract
Prolonged maintenance of therapeutically-relevant levels of broadly neutralizing antibodies (bnAbs) is necessary to enable passive immunization against infectious disease. Unfortunately, protection only lasts for as long as these bnAbs remain present at a sufficiently high concentration in the body. Poor pharmacokinetics and burdensome administration are two challenges that need to be addressed in order to make pre- and post-exposure prophylaxis with bnAbs feasible and effective. In this work, we develop a supramolecular hydrogel as an injectable, subcutaneous depot to encapsulate and deliver antibody drug cargo. This polymer-nanoparticle (PNP) hydrogel exhibits shear-thinning and self-healing properties that are required for an injectable drug delivery vehicle. In vitro drug release assays and diffusion measurements indicate that the PNP hydrogels prevent burst release and slow the release of encapsulated antibodies. Delivery of bnAbs against SARS-CoV-2 from PNP hydrogels is compared to standard routes of administration in a preclinical mouse model. We develop a multi-compartment model to understand the ability of these subcutaneous depot materials to modulate the pharmacokinetics of released antibodies; the model is extrapolated to explore the requirements needed for novel materials to successfully deliver relevant antibody therapeutics with different pharmacokinetic characteristics.
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SciScore for 10.1101/2022.05.24.493347: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Field Sample Permit: At regular intervals, aliquots of the supernatant were taken for measurement in the plate reader as described above, and the sampled volume was replaced with PBS. 2.6 In vitro diffusivity measurements: All fluorescence recovery after photobleaching (FRAP) experiments were conducted in the Stanford University Cell Sciences Imaging Facility (CSIF) at room temperature or 37 °C.
IACUC: 2.13 In vivo Centi-C10 mAb pharmacokinetic study: All animal studies were performed in accordance with National Institutes of Health guidelines and with the approval of the Stanford Administrative Panel on Laboratory Animal Care (APLAC-32109).Sex as a biological variable Female B6.Cg-Fcgrttm1Dcr … SciScore for 10.1101/2022.05.24.493347: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Field Sample Permit: At regular intervals, aliquots of the supernatant were taken for measurement in the plate reader as described above, and the sampled volume was replaced with PBS. 2.6 In vitro diffusivity measurements: All fluorescence recovery after photobleaching (FRAP) experiments were conducted in the Stanford University Cell Sciences Imaging Facility (CSIF) at room temperature or 37 °C.
IACUC: 2.13 In vivo Centi-C10 mAb pharmacokinetic study: All animal studies were performed in accordance with National Institutes of Health guidelines and with the approval of the Stanford Administrative Panel on Laboratory Animal Care (APLAC-32109).Sex as a biological variable Female B6.Cg-Fcgrttm1Dcr Prkdcscid Tg(FCGRT)32Dcr/DcrJ (The Jackson Laboratory, Stock No. 018441) mice age 12-14 weeks were administered Centi-C10 antibody via IV (retro-orbital) or SC injection under brief isoflurane anesthesia. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Fragment Goat Anti-Human IgG, Fcγ fragment specific (Jackson Immunoresearch, 109-036-008, RRID: AB_2337591), and incubating for 1 hr. Fragment Goat Anti-Human IgGdetected: (Jackson ImmunoResearch Labs Cat# 109-036-008, RRID:AB_2337591)Hydrogel samples were prepared for the in vivo study from a stock solution of 4 wt% HPMC-C12 dissolved in PBS containing the appropriate concentration of Centi-C10 antibody and 20 wt% NPs stock solution. Centi-C10suggested: NoneExperimental Models: Cell Lines Sentences Resources 2.9 Spike-pseudotyped lentivirus production and viral neutralization assays: Spike-pseudotyped lentivirus was produced in HEK293T cells as previously described. HEK293Tsuggested: RRID:CVCL_HA71)Spike-pseudotyped lentiviral neutralization assays were performed using HeLa cells overexpressing human ACE2, as described in. HeLasuggested: CLS Cat# 300194/p772_HeLa, RRID:CVCL_0030)46 HeLa/ACE2 cells were plated in 96-well clear bottom, white-walled plates 1 day prior to infection at a density of 5,000 cells per well. HeLa/ACE2suggested: JCRB Cat# JCRB1845, RRID:CVCL_B3LW)Experimental Models: Organisms/Strains Sentences Resources For in vitro release assays and diffusion measurements, a total concentration of 10 mg/mL rat IgG was loaded into the hydrogels (9 mg/mL rat IgG and 1 mg/mL fluorescently-labeled rat IgG). 2.3 Dynamic and flow rheometry: All rheometry experiments were performed on a torque-controlled Discovery Hybrid Rheometer (DHR2, TA Instruments). DHR2suggested: NoneFemale B6.Cg-Fcgrttm1Dcr Prkdcscid Tg(FCGRT)32Dcr/DcrJ (The Jackson Laboratory, Stock No. 018441) mice age 12-14 weeks were administered Centi-C10 antibody via IV (retro-orbital) or SC injection under brief isoflurane anesthesia. B6.Cg-Fcgrttm1Dcr Prkdcscid Tg(FCGRT)32Dcr/DcrJsuggested: RRID:IMSR_JAX:018441)Recombinant DNA Sentences Resources The library was subjected to 4 rounds of selection and ScFv were tested for the binding of SARS-CoV-2 RBD and blocking its interaction with of hu-ACE-2 on high-throughput surface plasmon resonance (SPR) on Carterra LSA Array SPR instrument (Carterra) equipped with HC200M sensor chip (Carterra) at 25 °C. 2.8 Antibody cloning into expression vectors for transient transfection: Heavy chain and light chain sequences of the complimentary determinant regions (CDRs) of centi-C10 ScFv was cloned into each constant region of human IgG1 heavy chain, and human kappa constant IGKC, in the mammalian expression pTT5 vector (Licensed from National Research Council of Canada, Toronto Canada), respectively. pTT5suggested: RRID:Addgene_52326)Cells were transfected using calcium phosphate transfection with 5 plasmids previously described in45 in the following ratios: 10 μg luciferase-containing lentivirus packaging vector (pHAGE-Luc), 3.4 μg FL SARS-CoV-2 Spike, 2.2 μg HDM-Tat1b, 2.2 μg pRC-CMV-Rev1b, and 2.2 μg HDM-Hgpm2. pHAGE-Lucsuggested: NonepRC-CMV-Rev1bsuggested: RRID:Addgene_164443)Software and Algorithms Sentences Resources 2.12 Pharmacokinetic (PK) modeling: The IV bolus data was fit to a two-phase exponential decay with the plateau constrained to zero in GraphPad Prism 9 to determine kelim; Vd was calculated from M0/C0, where C0 is the initial concentration from the two-phase exponential. GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)47 Simulated pharmacokinetic profiles were generated in MATLAB. MATLABsuggested: (MATLAB, RRID:SCR_001622)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
Results from scite Reference Check: We found no unreliable references.
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