Safety — Biological

Organisms and Use

• Escherichia coli K-12 (e.g., DH5α/XL1-Blue): non-pathogenic laboratory strains used only for cloning and plasmid propagation.
• Pichia pastoris (Komagataella phaffii): non-pathogenic yeast widely used in biotechnology for recombinant protein expression.
• Genetic parts: coding sequences for scFv, promoters, secretion signals; no virulence genes, toxins, or pathogenic elements.

Risk Identification

Identified risk: accidental contamination of cultures or improper handling.

Containment and Procedures

• Work conducted in BSL-1 facilities suitable for K-12 E. coli and P. pastoris.
• Culture handling only in designated areas; use biosafety cabinets when generating aerosols.
• Primary containment: safe pipetting, closed containers; secondary containment: restricted access, signage.

Mitigation and Disposal

• Mandatory PPE: lab coats, nitrile gloves, safety glasses; masks as needed.
• Autoclave all biological waste prior to disposal; incident reporting and logs maintained.
• Ongoing training in microbial handling and spill response.