Culture cells aseptically, count and passage them, control contamination, model growth, understand bioreactors and purification, and work under GMP with validated systems and batch data.

Cell Culture & Bioprocessing

Write a master batch record for "500 g of 10 percent w/w sugar solution": product name, a record number and version, materials (sugar 50.0 g, range 49.5 to 50.5 g; water 450 g, range 445 to 455 g), equipment (scale with an ID you assign), numbered steps, and fields for actual value, performed by and verified by.

Add a step to record the start time and end time, and a final step for appearance (clear, no undissolved sugar).

Issue yourself a copy: write "Batch 001" and the date on it.

Execute it: tare the scale, weigh the sugar and record the actual weight immediately in ink, then the water, then mix until dissolved. Record every value at the time.

Ask a friend or family member to watch the weighing steps and sign as verifier (or role-play both and note it).

Make one deliberate recording error and correct it with GDP: single line, correct value, initials, date, reason.

Write a planned deviation: for example, you mixed for 4 minutes instead of the written 2 minutes. Record what happened, the impact on the product and who you told.

Review your own completed record as QA would: any blank fields, missing signatures or unexplained changes? List them.

## Why it matters

Medicines, vaccines and many biological products must be made under **Good Manufacturing Practice (GMP)**: rules that make sure every batch is made the same validated way, by trained people, with a complete record. A patient cannot test their own medicine, so the record is the evidence. For many biological technicians, GMP is the daily working environment, and the basic skill is executing a batch record correctly. As the saying goes: if it is not documented, it did not happen.

## What GMP covers

GMP regulations and guidelines cover, among other things:

- **Personnel:** qualified, trained people with training records for every procedure they perform.
- **Facilities and equipment:** designed, qualified, cleaned and maintained (next lesson).
- **Materials:** approved suppliers, identity testing, quarantine until release, traceable lot numbers.
- **Procedures:** written, approved SOPs and batch records, followed exactly.
- **Records:** complete, accurate, contemporaneous, reviewed.
- **Quality unit:** an independent quality assurance (QA) function that approves procedures and releases or rejects batches.
- **Deviations, CAPA and change control:** anything unplanned is recorded and investigated; changes are assessed before they are made.

In the US, the core drug GMP regulations are 21 CFR Parts 210 and 211, with additional requirements for biologics in Parts 600 to 680. FDA's overview: https://www.fda.gov/drugs/pharmaceutical-quality-resources/current-good-manufacturing-practice-cgmp-regulations (Checked on 2026-10-23.) In the EU, EudraLex Volume 4 sets GMP: https://health.ec.europa.eu/medicinal-products/eudralex/eudralex-volume-4_en (Checked on 2026-10-23.) Other regions follow similar rules, often harmonised through PIC/S and ICH.

## The batch record

A **master batch record** is the approved template for making one batch of a product: materials and quantities, equipment, step-by-step instructions, process parameters and limits, sampling, and signature fields. For each batch, a controlled copy is issued and becomes the **executed batch record** as operators fill it in.

Executing a step typically means:

1. **Read** the step before doing it.
2. **Do** exactly what it says, with the equipment and materials it names.
3. **Record** the actual values (weights, times, temperatures, lot numbers, equipment IDs) at the time, in permanent ink or the electronic system.
4. **Sign and date** ("performed by").
5. A second trained person **verifies** critical steps (such as weighing or adding an ingredient) and signs "verified by".

## Good documentation practice (GDP)

GDP applies the ALCOA+ principles you learned in the lab data course:

- Write entries **at the time**, never from memory later, and never pre-fill.
- Record **actual** values with units, not "OK" or a tick where a number is required.
- Correct errors with a **single line**, the correct value, initials, date and a reason. No correction fluid, no overwriting.
- Never leave blanks: write "N/A" with initials if a field does not apply, as your procedure allows.
- Use your **own** signature or login; never sign for someone else.
- If something goes differently from the instruction, **stop** and record a **deviation** following your SOP, and tell your supervisor. Do not improvise and carry on.

**Worked example.** Step 12 says "Add 25.0 kg (24.8 to 25.2 kg) of buffer component X, lot to be recorded. Verify." You weigh 25.1 kg, record "25.1 kg", the lot number, the balance ID, the time, sign "performed by", and a colleague who watched the weighing signs "verified by". If you had weighed 25.4 kg, you would not just record it and continue: it is outside the range, so you stop and follow the deviation procedure.

## After the batch

QA reviews the executed batch record, along with test results, deviations and environmental data, before the batch can be **released**. Any gap (a missing signature, an unexplained correction) delays release and can trigger an investigation. You will review batch data yourself in this module's last lesson.

## Training reality

GMP training is given by the employer, specific to its procedures, and repeated regularly. Many community colleges and universities offer biomanufacturing certificates with GMP and aseptic modules. You do not work unsupervised in a GMP area until you are trained and qualified on that procedure.

## Common mistakes

- Filling in several steps at once at the end of the shift.
- Writing a value outside the range and carrying on.
- Ticking "verified" without watching the step.
- Using a stray piece of paper as a "rough record".
- Unsigned or undated corrections.

## Quick recap

- GMP: trained people, qualified equipment, controlled materials, written procedures, complete records, independent QA.
- Master batch record (template) vs executed batch record (this batch).
- Read, do, record actual values, sign, verify critical steps.
- GDP: contemporaneous, single-line corrections with reason, no blanks, no signing for others, stop and record deviations.

## Try it

Open the **Do** tab to write and execute a mock batch record for a kitchen-safe sugar solution.


Explain what GMP requires of people and records, and follow a batch record with good documentation practice.

Working under GMP

What is Good Manufacturing Practice GMP in Pharmaceuticals?

What is Batch Record Review? Explained in 2 minutes! | Free GMP Training video 2025

## Why it matters

GMP does not trust equipment or processes because they look right. It requires documented evidence that they work as intended, every time: this is **validation**. And for products that cannot be terminally sterilised, such as many biologics, the room itself is part of product protection: **cleanrooms** control particles and microbes, and people, the biggest source of contamination, enter them only after gowning in a fixed sequence. Technicians execute qualification protocols, perform environmental monitoring and gown many times a day.

## Equipment qualification: IQ, OQ, PQ

Before a new bioreactor, incubator, autoclave or HPLC is used for GMP work, it is qualified against its **user requirements specification (URS)**:

| Stage | Question | Examples |
|---|---|---|
| **DQ** design qualification | Is the design suitable for the URS? | Review of specifications and drawings |
| **IQ** installation qualification | Is it installed correctly, as specified? | Correct model and parts, utilities connected, calibration certificates, manuals and drawings filed, software version recorded |
| **OQ** operational qualification | Does it operate across its intended ranges? | Temperature control at low, mid and high set points; alarms trigger; door interlocks work |
| **PQ** performance qualification | Does it perform consistently under real conditions? | Temperature mapping of a loaded incubator over days; a sterilisation cycle with a real load and biological indicators |

Each stage has an approved **protocol** written before testing, with acceptance criteria. Results go in a **report**; any failure is a deviation that must be resolved. After qualification, equipment stays in a validated state through calibration, preventive maintenance, periodic review and **change control**: a change (new software version, moved equipment) is assessed and may need requalification.

## Process validation

Qualified equipment is necessary but not enough: the **process** must be shown to make product of the required quality consistently. FDA's process validation guidance describes three stages: **process design** (development defines critical parameters), **process qualification** (including process performance qualification batches at commercial scale) and **continued process verification** (ongoing monitoring of every batch). Cleaning procedures, analytical methods and computerised systems are validated too.

## Cleanrooms

Cleanrooms are classified by airborne particle counts. ISO 14644-1 defines classes such as ISO 5, 7 and 8. EU GMP Annex 1 (in EudraLex Volume 4: https://health.ec.europa.eu/medicinal-products/eudralex/eudralex-volume-4_en, checked on 2026-10-23) uses **grades A to D** for sterile manufacturing:

- **Grade A:** the critical zone where sterile product or open containers are exposed (inside a unidirectional airflow hood or isolator).
- **Grade B:** the background room around Grade A for aseptic work.
- **Grades C and D:** less critical steps, such as preparing solutions.

Key design features: HEPA-filtered air supply, many air changes per hour, a **pressure cascade** (cleaner rooms at higher pressure, so air flows outward toward dirtier areas), **airlocks** between grades, smooth cleanable surfaces, and controlled flows of people and materials.

**Environmental monitoring** checks that the room stays in control: particle counts, settle plates and air samples for microbes, contact plates on surfaces and on operators' gloves and gowns. Results are trended, and excursions are investigated. **Aseptic process simulations (media fills)** run the aseptic process with growth medium instead of product to show that operators and process do not introduce contamination.

## Gowning

People shed skin cells and microbes constantly, so gowning procedures are strict and sequential, moving from "dirty" to "clean":

1. Remove outdoor clothing and jewellery; wash hands.
2. Put on shoe covers or dedicated shoes, crossing a step-over bench.
3. Put on a hood or hair cover and face mask (and goggles as required).
4. Put on the sterile coverall without letting it touch the floor, then sterile boots over it.
5. Put on sterile gloves, often two pairs, sanitised regularly.
6. Check yourself in a mirror for gaps.

The order matters: you cover the head before the coverall, so hair and skin particles fall onto clothing you will cover, not onto the clean suit. Operators are **gowning qualified** (with contact plates taken from the gown) before entering Grade A/B areas, and requalified periodically. Inside, they move slowly and deliberately and never touch critical surfaces.

## Common mistakes

- Running OQ before IQ is approved, or testing without an approved protocol.
- Moving or modifying qualified equipment without change control.
- Letting the coverall touch the floor during gowning.
- Fast movements and talking over open product in Grade A.
- Treating environmental monitoring excursions as noise instead of trends to investigate.

## Quick recap

- IQ: installed correctly; OQ: operates across ranges; PQ: performs consistently in real use. Protocol first, report after.
- Validation covers processes, cleaning, methods and computerised systems; change control keeps the validated state.
- Cleanrooms: HEPA air, pressure cascade, airlocks; EU grades A to D; environmental monitoring and media fills.
- Gowning follows a fixed dirty-to-clean order; operators are gowning qualified.

## Try it

The **Do** tab has a quiz on qualification stages and gowning.


Explain equipment qualification (IQ, OQ, PQ), process validation, cleanroom grades and why gowning is done in a fixed order.

Validation and cleanrooms

Qualification Phases: IQ, OQ, and PQ Explained Simply

Aseptic Gowning for the Cleanroom

## Why it matters

Before a batch of drug substance can be released, someone has to confirm that every required test was done and every result meets its **specification**. A single missed out-of-specification (OOS) result can put an unsafe batch on the market; a sloppy investigation that "tests into compliance" can lead to regulatory action. Batch data increasingly lives in databases (LIMS and manufacturing execution systems), so being able to query it, not just read printouts, makes review faster and more reliable.

## Specifications, OOS and OOT

- A **specification** is the list of tests, methods and acceptance criteria a batch must meet, for example pH 5.8 to 6.4 or endotoxin not more than 0.5 EU/mg. Some limits are two-sided; others have only a lower limit (purity at least 95.0 percent) or only an upper limit (endotoxin).
- An **out-of-specification (OOS)** result falls outside an acceptance criterion.
- An **out-of-trend (OOT)** result is inside specification but unusual compared with previous batches, for example a sudden step change. It is a warning worth investigating, just like a Westgard warning in clinical QC.

Boundary values matter: if the limit is "not more than 0.5" then 0.5 passes and 0.51 fails. Specifications also state how results are rounded before comparison, so follow the written rule.

## What a batch data review checks

1. **Completeness:** every required test has a result, from a valid run, with the right method version.
2. **Compliance:** each result meets its specification.
3. **Trends:** results compared with previous batches (control charts).
4. **Related records:** the executed batch record is complete, deviations are closed or assessed, environmental monitoring is in control, and equipment was qualified and calibrated.
5. **Data integrity:** audit trails show no unexplained changes, deletions or repeated injections.

Only after QA is satisfied is the batch released, and a certificate of analysis (CoA) issued.

## Querying batch data in SQL

Suppose a LIMS has `results(batch_id, test, value, analyst)` and `specs(test, lower_limit, upper_limit, units)`, where a missing limit is stored as `NULL`. Joining them puts every result next to its limits:

```sql
SELECT r.batch_id, r.test, r.value, s.lower_limit, s.upper_limit
FROM results AS r
JOIN specs AS s ON s.test = r.test;
```

To keep only OOS results, add a condition that the value is below the lower limit **or** above the upper limit. In SQL, any comparison with `NULL` is "unknown" rather than true, so a test with no upper limit simply never fails the upper check, which is exactly what we want here. (If you write it the other way round, as "keep rows that are within limits", `NULL` limits would wrongly exclude rows, so be deliberate.)

**Worked example.** pH limits 5.8 to 6.4. Batch B-202 has pH 6.6: 6.6 > 6.4, so it is OOS. Batch B-204 has pH exactly 6.4: not greater than 6.4, so it passes.

## What happens after an OOS

In the US, FDA's guidance *Investigating Out-of-Specification (OOS) Test Results for Pharmaceutical Production* sets expectations: https://www.fda.gov/regulatory-information/search-fda-guidance-documents/investigating-out-specification-oos-test-results-pharmaceutical-production-level-2-revision (Checked on 2026-10-23.) In outline:

1. **Phase I, laboratory investigation:** the analyst and supervisor promptly check for an obvious, documented laboratory error (wrong dilution, instrument failure, calculation error), preserving the original samples and data. Only a clearly identified lab error can invalidate the result.
2. **Phase II, full-scale investigation:** if no lab error is found, the investigation extends to manufacturing: batch record, deviations, raw materials, equipment and other batches that might be affected.
3. **Retesting and resampling** follow predefined rules. Testing again and again until a passing result appears ("testing into compliance") is not acceptable, and averaging cannot be used to hide an individual OOS value.
4. The conclusion, root cause and CAPA are documented, and QA decides the batch's fate.

## Common mistakes

- Reviewing only the tests that look interesting, not checking completeness.
- Treating a value exactly at the limit as a failure (or a value just past it as a pass) without reading the specification wording.
- Mishandling `NULL` limits in queries, which silently drops or wrongly flags rows.
- Discarding an OOS result as "probably a lab error" without evidence.
- Ignoring OOT results because they are technically in specification.

## Quick recap

- Specifications define tests and acceptance criteria; OOS is outside them, OOT is unusual but inside.
- Review covers completeness, compliance, trends, related records and data integrity before QA release.
- In SQL, join results to specs and filter value < lower OR value > upper; know how `NULL` behaves.
- OOS investigations: Phase I lab check, Phase II full-scale, controlled retesting, documented conclusion; never test into compliance.

## Try it

Open the **Do** tab and write the `WHERE` clause that flags out-of-specification results.


Review batch test results against specifications, flag out-of-specification results in SQL and know what happens next.

Batch data review

Investigation of Out of Specification Results | OOS Investigation

Batch Record Review Process in Pharmaceutical Manufacturing. GMP Batch Release and Disposition.

Follow a batch record correctly, explain IQ/OQ/PQ and cleanroom gowning, and flag out-of-specification batch results in SQL.