I need to write Cleaning Validation Protocol for cleaning validation of equipment and working surface. The method, which I have to use, is determination of residue protein by Bradford protein assay. In this context I have some questions and I would be very grateful if someone could help me:
- The number of samples should be three, but how I can to determine the time between taking samples (5 min, 10min, 30 min or 1h etc)? How long after cleaning should begin sampling?
- How I can to determine the holding time between the end of working process and cleaning?
- Where can I find template for Cleaning Validation Protocol by using Bradford protein assay?
Thank you in advance for your help.
[b][COLOR=“darkred”]You must take a sample normally after the surface dries in your case. May be 15 to 20 minits.
Take samples at 5 min, 10 min, 15min, 30 min and 1 hour.Plot a graph to show its cleanability.
Make sure that the assay is with in the snsitivity range to detect the protein.
In your case the holding time is just 15 to 20 minits. The reason is you are not finding any Bioburden in this study. You are finding the protein residues after cleaning on a Biopharmaceutical manufacturing process.
Your intention might be:
How effectively the equipment is cleaned?
How effectively the protein is removed to start another product if you work on campgain basis?
Limits of detectection of previous product carry overs.
Yes, there are some published papers on these things which I found in year 2003 & 2004
I will certainly attach those references here soon. I need some time to find them.[/color][/b]
Development of surface swabbing procedures for a cleaning validation program in a biopharmaceutical manufacturing facility
Silvia Lombardo, Prasad Inampudi, Anthony Scotton, Gary Ruezinsky, Randall Rupp, Somesh Nigam
Biotechnology and Bioengineering
Volume 48, Issue 5, pages 513–519, 5 December 1995
An Integrated Approach for Validating Cleaning Procedures in Biopharmaceutical Manufacturing Facilities
. INAMPUDI, S. LOMBARDO,†, G. RUEZINSKY, T. BALTRUS, J. DUGGER, P. REMSEN, R. RUPP, S. NIGAM
Article first published online: 17 DEC 2006
With reference to your queries,
- The number of samples should be three, but how I can to determine the time between taking samples (5 min, 10min, 30 min or 1h etc)?
How long after cleaning should begin sampling?
= I am in agreement with Mr. Durga by Logic as explained above.
How I can to determine the holding time between the end of working process and cleaning?
= Now the role of DEHT study starts & you have to design the study based on the product matrix alongwith risk assessment. Basically this study is being done to have a assurance on degredation of product nature.
Where can I find template for Cleaning Validation Protocol by using Bradford protein assay?
= Leave a message from your mail to below mentioned ID.
Happy Reading !
Thank you for your answers, guys! You gave me very useful information.
The aim of this study is to determine the efficiency of the current cleaning procedure.
From my point of view the time in which establishes a maximum adsorption of proteins is the time in which cleaning is most efficient. So that the cleaning procedure should be perform in different time after marking of the surface/equipment with protein solution. For example 5 min after marking with protein solution, 10 min and etc. In my opinion the protein concentration in samples, taken after cleaning in later time (for example 30 min after marking), must be lower. This could prove that the cleaning procedure is most efficient right now after the end of working process.
Can you tell me if these thoughts are correct?
Thank you in advance for your time and help.
You must polt the graphs
5minits- Protein concentration
10 minits- Protein concentration
15 minits -Protein concentration
20-Minits -protein concentration
25 minits -protein cocentration
1 hour- protein concentration
If you look the graph you will find out that at certain point there is a very high protein concentration as per the assay. This is the point time in minits where maximum protein is desorped from the surface.The later concentrations show decline in concentrations.At a particular time you will find the concentrates of these residues below the levels prescribed by the regulatory authorities.
You must atleast carry our 5 to six cleaning cycles and map graphs.
What you get from those Graphs?
These graphs gives you a trend analysis too which is very important when you qualify and quantify your cleaning methodologies on Risk methodology.
From the graphs you also evaluate maximum and minimum cleaning times so that the whole process line is clear from protein residues.
From those you can also have alert limits, action limits and a validated ranges of concentrations found after cleaning.
Your approach is very right. I just added a direction to your approach.
What is the Microbial Limit for swabs for non-sterile area. We manufacture sterile serum and we clean the non-sterile equipment e.g filter housings, hoses tanks using CIP. Our current limit for Bioburden is ≤ 20cfu/ml which I think is quiet low for a non-sterile area.
I use sterile swabs and swab 4x4cm area and collect the swab in to 5mL saline solution.
Could you also give references to GMP guidelines.
Thanks in advance