# Cryopreservation of Anopheles Embryos

> **NIH NIH R44** · SANARIA, INC. · 2021 · $962,110

## Abstract

ABSTRACT: PfSPZ Vaccine is currently on track to be the world’s first licensed vaccine against malaria and
the first vaccine against a human parasitic disease. PfSPZ Vaccine is composed of aseptic, attenuated,
metabolically active, purified, vialed and cryopreserved Plasmodium falciparum (Pf) sporozoites (SPZ).
Manufacture of PfSPZ Vaccine shares all steps (except radiation attenuation) with PfSPZ Challenge that is
used by researchers worldwide in controlled human malaria infections to evaluate experimental malaria drugs,
candidate vaccines, and natural immunity or resistance to malaria. The third product under clinical testing is
PfSPZ-CVac, PfSPZ Challenge given under chemoprophylactic cover. PfSPZ Vaccine has protected against
homologous (same strain) and heterologous (different strains) of Plasmodium falciparum (Pf) malaria in clinical
trials conducted in the U.S., Germany, Tanzania, and Mali. To date, 80–100% protection lasting for at least 8
months against heterologous and 14 months against homologous challenge infections has been demonstrated.
Ongoing trials in Gabon, Mali, and Equatorial Guinea are testing optimized regimens in preparation for Phase 3
clinical trials starting in 2020 in support of a Biologics License Application (BLA) to the FDA in 2021. All PfSPZ
products are produced in mosquitoes reared aseptically from disinfected eggs that are harvested from a
continuously-maintained Anopheles stephensi colony. The goals of this project are to submit a Biologics
Master File for Sanaria’s cryopreserved A. stephensi SDA500 strain in support of the manufacture of PfSPZ-
based products and to manufacture one lot of PfSPZ Challenge using mosquitoes derived from cryopreserved
eggs, and also to adapt the methodology for large scale cryopreservation of A. gambiae. Fundamental to this
process is the elevation of mosquito production from banks of cryopreserved eggs at full cGMP compliance.
The work will involve cryopreservation of eggs at hugely increased scale and the generation of a master egg
bank (MEB) and working egg bank (WEB) of cryopreserved A. stephensi SDA500 eggs. The work is
incorporated into three specific aims: 1) Complete the GMP-compliant manufacture of a MEB and WEB which
will entail development of novel release and stability assays, generation of SOPs and Certificates of Analysis,
establishment of a stability program for cryopreserved eggs and submission of a Biologics Master File to the
FDA; 2) Manufacture, using mosquitoes derived from the cryopreserved WEB, one lot of PfSPZ Challenge for
use in clinical trials; and 3) Cryopreserve A. gambiae eggs at scale to support mosquito release programs.
Cryopreservation of Anopheles eggs is a breakthrough enabling technology that will be available to support
Anopheles mosquito research, particularly in laboratories that need to retain genetically modified strains and
lines but have limited resources to do so.

## Key facts

- **NIH application ID:** 10109966
- **Project number:** 5R44AI088853-07
- **Recipient organization:** SANARIA, INC.
- **Principal Investigator:** Peter F. Billingsley
- **Activity code:** R44 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $962,110
- **Award type:** 5
- **Project period:** 2010-08-01 → 2023-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10109966

## Citation

> US National Institutes of Health, RePORTER application 10109966, Cryopreservation of Anopheles Embryos (5R44AI088853-07). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10109966. Licensed CC0.

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